Unraveling electronic absorption spectra using nuclear quantum effects: Photoactive yellow protein and green fluorescent protein chromophores in water.
Many physical phenomena must be counted to accurately model the shapes of the optical spectral line of the phase phase phase phase solution, from the sampling of the chromophore solvent configurations to the electronic-vibrational transitions leading to the structure. Vibronic thin. Here, here we explore here in depth the role of nuclear quantum effects, direct and indirect solvent effects and vibronic effects in the calculation of the optical spectrum of solveed anionic chromophores of green fluorescent proteins and yellow photoactive proteins. By analyzing the chromophore and solvent configurations, vertical excitement energy distributions, the absorption spectra calculated in the overall approach and the absorption spectra calculated throughout a Frank Thermal at zero temperature, we show how solvents, nuclear quantum effects, and vibronic transitions modify optical absorption spectra.
We find that, including nuclear quantum effects in the sampling of chromophore solvent configurations using the integrated molecular ab initio hath simulation causes an improvement of spectral forms through three mechanisms. The three mechanisms that lead to the widening of the line form and a better description of the high-energy tail result in a softening of heavy atom bonds in the chromophore that couples with optically shiny state, expanding the distribution of energies. Vertical excitement of plus diverse solvent environments, and redistribution of the spectral weight of the vibrone transition 0-0 to vibronic transitions of higher energy during the computer of the Franck conton spectrum in a frozen solvent pocket.
The absorption spectra calculated using the combined assembly together as well as the zero-thement temperature, the zero temperature condo approach gave significant improvements in the spectral shape and width with respect to calculated spectra with The overall approach. The use of the approach combined with configurations sampled from integrated trajectory the molecular dynamics of the trajectories has a significant step forward in the precise modeling of the absorption spectra of the aquately solvated chromophores.
Cipergenesis, a mutagenesis approach that produces small random permuted traffic protein libraries in a targeted region: Test on the green fluorescent protein.
The permuted circular proteins (CPP) represent a new type of mutant proteins with covalently bonded originne termini through a peptide connector and open to any other place of the polypeptide backbone to create new ends. CPPs find broad biotechnology applications because their properties can be very different from those of parental protein. However, the actual challenge of creating successful CPP is to identify these peptide bonds that can be broken to create new terminals and to ensure functional and well-folded CPPs. In this document, we describe Cipergenesis, a combinatorial mutagenesis approach that uses two oligonucleotide libraries to amplify a Circular PCR gene, start and end with a targeted target region.
This approach creates small transmitted traffic genes libraries easily cloned in the right direction and the right frame using two different restriction sites encoded in oligonucleotides. Once expressed, protein libraries have a single sequence diversity, comprising CPPs that have ordinary breakpoints between adjacent amino acids located in the target area as well as CPCs with new user-defined truncations and Repetitions of some amino acids. Cipergenesis has been tested at the G134-H148 lid region of the Green Fluorescent Protein (GFP), revealing that the most fluorescent variants were those beginning to LU141 and ending with amino acids Tyr145, Tyr143, GLU142, Leu141, Lys140 and H139.
The purification and biochemical characterization of certain variants suggested a differential expression, a magnitude of the solubility and the maturation of the mutant proteins, because the probable cause of the variability of the fluorescence intensity observed in the colonies.
Unraveling electronic absorption spectra using nuclear quantum effects: Photoactive yellow protein and green fluorescent protein chromophores in water.
Structural Insight on the photochemistry of green fluorescent proteins divided: a unique role for label label.
Oligohistidine affinity labels (its tags) are commonly fused to proteins to facilitate their purification via a chromatography on affinity metal. These tags are usually assumed to have a minimum impact on the properties of the melting protein because they have no propensity to form controlled elements and are small enough not to significantly affect solubility or size. Here, we signal structures of two variants of the truncated green fluorescent protein (GFP), that is to say a fractionated GFP with a removable strand β, which have been found to behave differently in the presence of light.
In these structures, the N-terminal its tag and several neighboring residues play a highly unusual structural and functional role in the stabilization of the truncated GFP by substituting as a substitution substitution substitution in the throat released by the native strand. This discovery provides an explanation of the binding and photodissociation properties of peptides apparently very different from the fractional proteins involving β-strands 10 and 11.
Description: The Nrf2 antioxidant response pathway plays an important role in the cellular antioxidant defense. Nrf2, a basic leucine zipper transcription factor, induces the expression of antioxidant and phase II enzymes by binding to the ARE (antioxidant response element) region of the gene promoter. Under basal conditions, Nrf2 is retained in the cytosol by binding to the cytoskeletal protein Keap1. Upon exposure to oxidative stress or other ARE activators, Nrf2 is released from Keap1 and translocates to the nucleus, where it can bind to the ARE, leading to the expression of antioxidant and phase II enzymes that protect the cell from oxidative damage. The ARE Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by ARE located upstream of the minimal TATA promoter. After transduction, activation of the Nrf2 antioxidant response pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The SRE (Serum Response Element) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the Serum Response Element located upstream of the minimal TATA promoter . After transduction, activation of the MAPK/ERK signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The Myc Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the Myc response element located upstream of the minimal TATA promoter and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the Myc signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The UAS (Upstream Activation Sequence) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by a multimerized GAL4 upstream activation sequence (UAS) located upstream of the minimal TATA promoter and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the UAS-controlled signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The p53 Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by p53 response elements located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, p53-regulated gene expression in the target cells can be monitored by measuring the luciferase activity.
Description: The Hypoxia Response Element (HRE) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by four copies of a hypoxia response elements (HRE) located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the induction of hypoxia in the target cells can be monitored by measuring the luciferase activity.
Description: The Hippo pathway regulates cell proliferation and cell death. It is activated by high cell density and cell stress to stop cell proliferation and induce apoptosis. The mammalian Hippo pathway comprises MST kinases and LATS kinases. When the Hippo pathway is activated, MST kinases phosphorylate LATS kinases, which phosphorylate transcriptional co-activators YAP and TAZ. Unphosphorylated YAP and TAZ remain in nucleus and interact with TEAD/TEF transcriptional factors to turn on cell cycle-promoting gene transcription. However, when phosphorylated, YAP and TAZ are recruited from the nucleus to the cytosol, so that the YAP and TAZ-dependent gene transcription is turned off. Dysfunction of the Hippo pathway is frequently detected in human cancer and its down-regulation correlates with the aggressive properties of cancer cells and poor prognosis. The TEAD Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the TEAD response elements located upstream of the minimal TATA promoter. After transduction, activation of the Hippo pathway in the target cells can be monitored by measuring the luciferase activity._x000D_
Description: The STAT3 Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene under the control of STAT3-responsive element located upstream of the minimal TATA promoter. After transduction, activation of the STAT3 signaling pathway in the target cells can be monitored by measuring the luciferase activity._x000D_
Description: The STAT5 Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene under the control of STAT5-responsive element located upstream of the minimal TATA promoter. After transduction, activation of the STAT5 signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The NF-κB Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by four copies of the NF-κB response element located upstream of the minimal TATA promoter. After transduction, activation of the NF-κB signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The main role of the cAMP response element, or CRE, is mediating the effects of Protein Kinase A (PKA) by way of transcription. Upon phosphorylation, CREB forms a functionally active dimer that binds the CRE element within the promoters of target genes and activates transcription. CRE is at the focus of many extracellular and intracellular signaling pathways, including cAMP, calcium, GPCR (G-protein coupled receptors) and neurotrophins. The cAMP/PKA signaling pathway is critical to numerous life processes in living organisms.The CRE/CREB Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by multimerized cAMP response element (CRE) located upstream of the minimal TATA promoter. After transduction, activation of the cAMP/PKA signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The NFAT Luciferase-RFP Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase and RFP (Red Fluorescent Protein) cassette driven by the NFAT response element located upstream of the minimal TATA promoter and a hygromycin or puromycin selection gene to generate stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity or RFP expression. RFP fluoresces red-orange when excited; it has an excitation wavelength of 553 nm, and an emission wavelength of 574 nm.
Description: The NFAT Luciferase-RFP Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase and RFP (Red Fluorescent Protein) cassette driven by the NFAT response element located upstream of the minimal TATA promoter and a hygromycin or puromycin selection gene to generate stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity or RFP expression. RFP fluoresces red-orange when excited; it has an excitation wavelength of 553 nm, and an emission wavelength of 574 nm.
Description: The IL-2 Promoter Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the human IL-2 promoter. After transduction, activation of the IL-2 signaling pathway in the target cells can be monitored by measuring the luciferase activity._x000D_
Description: The IL-8 Promoter Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the human IL-8 promoter. After transduction, activation of the IL-8 signaling pathway in the target cells can be monitored by measuring the luciferase activity._x000D_
Bald Lentiviral Pseudovirion (Luciferase Reporter)
Description: The bald lentiviral pseudovirion was produced without envelope glycoproteins such as VSV-G or SARS-CoV-2 spike. It contains the firefly luciferase gene driven by a CMV promoter as the reporter. The bald lentiviral pseudovirion can serve as a negative control when studying virus entry initiated by specific interactions between virus particles and receptors._x000D_
Description: The NFAT Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the NFAT response element located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (hygromycin, puromycin, or G418) for the selection of stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The NFAT Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the NFAT response element located upstream of the minimal TATA promoter and an antibiotic selection gene (hygromycin or puromycin) for the selection of stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The NFAT Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by the NFAT response element located upstream of the minimal TATA promoter and an antibiotic selection gene (hygromycin or puromycin) for the selection of stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The Xenobiotic response element (XRE) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by three copies of an XRE located upstream of the minimal TATA promoter (Figure 1), and an antibiotic selection gene (puromycin) for the selection of stable clones. After transduction, the activation of aryl hydrocarbon receptor (AhR) in the target cells can be monitored by measuring the luciferase activity.
Description: The SBE Luciferase Reporter Lentivirus (TGFβ/SMAD signaling pathway) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by multimerized SBE-responsive element located upstream of the minimal TATA promoter. After transduction, activation of the TGFβ/SMAD signaling pathway can be monitored by measuring the luciferase activity._x000D_
Description: The NFAT Luciferase-eGFP Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase and eGFP cassette driven by the NFAT response element located upstream of the minimal TATA promoter (Figure 1) and a puromycin selection gene to generate stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity or eGFP expression.
Description: The NFAT Luciferase-eGFP Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase and eGFP cassette driven by the NFAT response element located upstream of the minimal TATA promoter (Figure 1) and a puromycin selection gene to generate stable clones. After transduction, activation of the NFAT signaling pathway in the target cells can be monitored by measuring the luciferase activity or eGFP expression.
Description: The bald VSV Delta G (Luciferase Reporter) was produced without envelope glycoproteins. It contains the firefly luciferase gene as the reporter. The bald VSV Delta G (Luciferase Reporter) can serve as a negative control when studying virus entry initiated by specific interactions between virus particles and receptors.
Description: The bald VSV Delta G (Luciferase Reporter) was produced without envelope glycoproteins. It contains the firefly luciferase gene as the reporter. The bald VSV Delta G (Luciferase Reporter) can serve as a negative control when studying virus entry initiated by specific interactions between virus particles and receptors.
Description: The stress-activated protein kinase/c-jun N-terminal kinase (SAPK/JNK) family of proteins includes mitogen-activated protein kinases (MAPKs) that are activated by stress, inflammatory cytokines, mitogens, oncogenes, and inducers of cell differentiation and morphogenesis. Upon activation of the SAPK/JNK pathway, MAP Kinase Kinases phosphorylate and activate JNKs. The activated JNKs translocate to the nucleus where they phosphorylate and activate transcription factors such as c-Jun. c-Jun then binds to the activator protein-1 (AP1) response element and induces AP1 transcription. The AP1 Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by AP1 response element located upstream of the minimal TATA promoter. After transduction, activation of the JNK signaling pathway and AP1 mediated activity in the target cells can be monitored by measuring the luciferase activity.
Description: The NFAT reporter (Luciferase)-THP-1 cell line is designed for monitoring the NFAT (nuclear factor of activated T-cells) signaling pathway in THP-1 cells by measuring luciferase activity. It contains a firefly luciferase gene driven by the NFAT response element located upstream of the minimal TATA promoter. Upon activation by NFAT activators such as Ionomycin, endogenous NFAT transcription factors bind to the DNA response elements, inducing transcription of the luciferase reporter gene.
GAS Luciferase Reporter Lentivirus (IFN-γ/JAK/STAT1 Pathway)
Description: The GAS Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by three copies of the interferon gamma (IFN-γ) activated sites (GAS) located upstream of the minimal TATA promoter and a puromycin selection gene for the selection of stable clones. After transduction, the GAS-regulated gene expression in the target cells can be monitored by measuring the luciferase activity.
Description: The STAT3 Luciferase Reporter THP-1 cell line is designed for monitoring the STAT3 signal transduction pathway. It contains a firefly luciferase gene driven by STAT3 response elements located upstream of the minimal TATA promoter. After activation by cytokines or growth factors, endogenous STAT3 binds to the DNA response elements, inducing transcription of the luciferase reporter gene.
Description: Human IL-2 reporter construct is stably integrated into the genome of Jurkat T-cells. The firefly luciferase gene is controlled by a human IL-2 promoter.
Description: The JAK/STAT pathway is activated by various cytokines and growth factors and plays a critical role in cell growth, hematopoiesis, and immune response. In mammals, there are four JAKs (JAK1, JAK2, JAK3 and TYK2) and seven STAT proteins. IFNα is a Type I interferon. Binding of IFNα to its receptor leads to the activation of JAK1 and TYK2, which in turn phosphorylate and activate STAT1 and STAT2. The phosphorylated STAT1 and 2 form a heterodimer and bind to IRF9/p48, forming a protein complex ISGF3. This complex translocates to the nucleus and binds to the ISRE (Interferon Stimulated Response Element) in the promoter region, thereby promoting transcription of interferon-inducible genes. The ISRE Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by multimerized ISRE response element located upstream of the minimal TATA promoter. After transduction, the activity of Type I interferon-induced JAK/STAT signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Description: The SARS-CoV-2 Spike Pseudotyped Lentivirus were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1) as the envelope glycoproteins instead of the commonly used VSV-G. These pseudovirions also contain the firefly luciferase gene driven by a CMV promoter, therefore, the spike-mediated cell entry can be conveniently measured via luciferase reporter activity. The SARS-CoV-2 Spike pseudotyped lentivirus can be used to measure the activity of neutralizing antibody against SARS-CoV-2 in a Biosafety Level 2 facility._x000D_ _x000D_
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection._x000D_The SARS-CoV-2 Spike Pseudotyped Lentivirus were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1) as the envelope glycoproteins instead of the commonly used VSV-G. These pseudovirions also contain the firefly luciferase gene driven by a CMV promoter, therefore, the spike-mediated cell entry can be conveniently measured via luciferase reporter activity. The SARS-CoV-2 Spike pseudotyped lentivirus can be used to measure the activity of neutralizing antibody against SARS-CoV-2 in a Biosafety Level 2 facility._x000D_ _x000D_
Description: Recombinant HEK293 cells expressing the firefly luciferase gene under the control of cAMP response element (CRE), and with forced expression of human GIPR (Gastric Inhibitory Polypeptide receptor; NM_000164.4). Activation of GIPR in these cells can be monitored by measuring luciferase activity._x000D_The functionality of the GIPR/CRE Luciferase Reporter HEK293 Cell Line was validated in a dose-response assay using agonists gastric inhibitory peptide (GIP) and tirzepatide hydrochloride. These agonists induce luciferase activity in a dose-dependent manner as depicted in Figure 1._x000D_
_x000D_Figure 1. Illustration of the GIPR/CRE Luciferase Reporter HEK293 Cell line.
Description: Recombinant HEK293 cell line stably expressing full-length human Calcitonin receptor-like receptor (CALCRL/CRLR/CLR; accession number: NM_005795) and containing a firefly luciferase gene under the control of multimerized cAMP response element (CRE). This cell line can be used to measure the EC50 and IC50 of CALCRL agonists and antagonists using the luciferase reporter activity.
Description: The Glucocorticoid Receptor (GR)-GAL4 Luciferase Reporter Jurkat Cell Line contains an engineered transcription factor stably integrated into the genome of Jurkat cells, which consists of the glucocorticoid receptor ligand binding domain fused to the DNA binding domain of GAL4. This fusion construct activates firefly luciferase expression which is under the control of a multimerized GAL4 upstream activation sequence (UAS). This allows for specific detection of glucocorticoid-induced activation of the glucocorticoid receptor without the need for individual transcriptional targets and with low cross-reactivity for other nuclear receptor pathways. This cell line is validated for response to stimulation of dexamethasone and to the treatment with mifepristone, an inhibitor of the glucocorticoid signaling pathway.
Description: The ADAR1 Luciferase Reporter HEK293 cell line is designed to monitor RNA editing by Adenosine deaminase acting on RNA (ADAR1). This cell line stably expresses ADAR1 under the control of a CMV promoter and a separate ADAR editing reporter construct expressed under the control of another CMV promoter. The reporter contains the gene encoding firefly luciferase, which is constitutively expressed in the cells, upstream of the gene encoding the GluA2 ADAR substrate followed by the Renilla luciferase gene. The sequence corresponding to GluA2 has been modified to contain an amber stop codon (UAG). When edited by ADAR, this stop codon (UAG) will be changed to UIG (A to I edit), which is read as tryptophan (UGG) by the translation machinery. This edit allows translation to occur all the way to the end of the reporter mRNA and results in the expression of Renilla luciferase. Conversely, in the absence of ADAR1 activity, translation terminates at the stop codon and Renilla is not expressed. Reporter activity is read out as the Renilla Luciferase/Firefly luciferase ratio whereby inhibition of ADAR activity, and thus the UAG (stop) to UGG (tryptophan) conversion rate, will result in a dose-dependent decrease in the Renilla luciferase/Firefly luciferase ratio.
Minicircle Single Reporter: CMV-Luciferase Parental Plasmid (10 µg)
Description: Recombinant Jurkat cell line expressing firefly luciferase under the control of an NFAT response element, and with constitutive expression of human TMIGD2 (Transmembrane and immunoglobulin domain containing 2; CD28H; NM_144615). Expression of the firefly luciferase gene is driven by NFAT response elements located upstream of the minimal TATA promoter. Activation of the NFAT signaling pathway in these cells can be monitored by measuring luciferase activity.
Description: This recombinant Jurkat cell line is a biologically relevant system to measure activation of the IL-15 cytokine receptor by IL-15. The cells were engineered for constitutive expression of human CD122 (IL-15Rβ; IL-2Rβ; NM_000878.4), and conditional expression of firefly luciferase driven by STAT5 response elements located upstream of the minimal TATA promoter. Expression of CD122 allows formation of a functional IL-15 receptor at the surface of Jurkat cells, which naturally express high levels of CD132 (also known as IL-15 receptor subunit γc). Activation of the STAT5 signaling pathway in response to IL-15 or IL-15 analogs can be monitored by measuring luciferase activity.
Description: Recombinant Jurkat T cells expressing both firefly luciferase and enhanced GFP (eGFP) under the control of NFAT response elements located upstream of the minimal TATA promoter. Activation of the NFAT signaling pathway can be monitored by examining either firefly luciferase or eGFP expression.
Description: Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems. Fast and convenient. Homogeneous "mix-and-measure" assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 2 min. Kit size: 1000 tests. Detection limit: 2 fg luciferase.
Description: Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems. Fast and convenient. Homogeneous "mix-and-measure" assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 2 min. Kit size: 1000 tests. Detection limit: 2 fg luciferase.
Description: Bright bioluminescent reagent system for rapid quantitation of luciferase reporter gene expression in transfected cells and high-throughput drug screens. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase and as few as 4 cells. Plus, the emitted light is linear over seven orders of magnitude. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, on LJL Analyst, Berthold Luminometer, Top-Count, MicroBeta counters, chemiluminescent image plate readers (CLIPR/LeadSeeker). Assay reagents compatible with all liquid handling systems. Fast and convenient. Homogeneous "mix-and-measure" assay allows detection of luciferase levels within 10 minutes. The optimally combined reagent system allows a single addition step, and simultaneous cell lysis and detection. Robust and amenable to HTS. Z factors of 0.6 to 0.8 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 2 min. Kit size: 1000 tests. Detection limit: 2 fg luciferase.
Description: Recombinant Jurkat cell line expressing firefly luciferase under the control of an IL-2-responsive promoter, and with constitutive expression of human KIR3DL3 (Killer Cell Immunoglobulin Like Receptor, Three Ig Domains and Long Cytoplasmic Tail 3; GenBank accession #BC143802.1 corresponding to KIR3DL3*00402 allele). HHLA2 (B7-H7) mediates an immune-stimulatory signal via TMIGD2 (Transmembrane and immunoglobulin domain containing 2; CD28H) in naïve T cells while it delivers an immune-inhibitory signal through KIR3DL3 in activated T cells and Natural Killer (NK) cells.
VSV-G Pseudotyped VSV Delta G (Luciferase Reporter)
Description: The VSV-G Pseudotyped VSV Delta G (Luciferase Reporter) was produced by re-expression of VSV-G as the envelope glycoprotein using the VSV Delta G system in which VSV-G is deleted. The pseudovirions contain the firefly luciferase gene; therefore, the VSV-G mediated cell entry can be measured via luciferase activity. The VSV-G Pseudotyped VSV Delta G (Luciferase Reporter) can be used as a positive control of transduction for other VSV pseudotypes containing the envelope glycoproteins of heterologous viruses in a Biosafety Level 2 facility.
VSV-G Pseudotyped VSV Delta G (Luciferase Reporter)
Description: The VSV-G Pseudotyped VSV Delta G (Luciferase Reporter) was produced by re-expression of VSV-G as the envelope glycoprotein using the VSV Delta G system in which VSV-G is deleted. The pseudovirions contain the firefly luciferase gene; therefore, the VSV-G mediated cell entry can be measured via luciferase activity. The VSV-G Pseudotyped VSV Delta G (Luciferase Reporter) can be used as a positive control of transduction for other VSV pseudotypes containing the envelope glycoproteins of heterologous viruses in a Biosafety Level 2 facility.
Description: The TCF/LEF Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene under the control of TCF/LEF-responsive element located upstream of the minimal TATA promoter. After transduction, activation of the Wnt/β-catenin signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Hippo Pathway TEAD Luciferase Reporter MCF7 cell line
Description: The TEAD Reporter - MCF7 cell line contains the firefly luciferase gene under the control of TEAD responsive elements stably integrated into the human breast cancer cell line, MCF7. Inside the cells, basal unphosphorylated YAP/TAZ remains in the nucleus and induces the constitutive expression of luciferase reporter. The cell line is validated for the inhibition of the expression of luciferase reporter by the activators of the Hippo pathway.
Description: Bioluminescent reagent system for rapid quantitation of firelfy and Ranilla luciferase reporter gene expression in transfected cells. Key Features: High sensitivity and wide detection range. Detection of as little of 2 fg luciferase. Compatible with routine laboratory and HTS formats. Assays can be performed in tubes or microplates, and measured with any luminometer. Can be readily automated on HTS liquid handling systems. Fast and convenient. Three step assay allows detection of dual luciferase levels within 20 minutes. Method: Luminescence. Samples: Cells etc. Species: All. Procedure: Assay takes 20 min. Kit size: 100 tests. Detection limit: 2 fg luciferase.
B2M Knockout NFAT Luciferase Reporter Jurkat Cell Line
Description: B2M (Beta-2-Microglobulin) has been genetically removed by CRISPR/Cas9 genome editing from NFAT Luciferase Reporter Jurkat cells. Expression of the firefly luciferase gene is driven by NFAT response elements located upstream of the minimal TATA promoter. Activation of the NFAT signaling pathway in these cells can be monitored by measuring luciferase activity.
Description: The anti-BCMA CAR Jurkat/NFAT-luciferase reporter cell line is a stable cell line made from the anti-BCMA scFV CAR lentivirus (BPS Bioscience #79701). It has been validated for anti BCMA-CAR expression by FACS, and for functional activation stimulated by both soluble BCMA protein (BPS Bioscience #79467) and BCMA/CHO target cells (BPS Bioscience #79500).
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and human ACE2 may offer protection against the viral infection. Numerous SARS-CoV-2 variants have been identified so far. These variants contain a number of mutations that may increase morbidity and mortality and allow the virus to spread more easily and quickly than the original strain.BPS Bioscience has launched a series of Spike Variants (SARS-CoV-2) Pseudotyped Lentivirus (Luc reporter). The Spike (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike Variant (see below for mutation details) as the envelope glycoproteins instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter, therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike Variants (SARS-CoV-2) Pseudotyped Lentivirus Pack (Luciferase Reporter) contains a collection of 12 Spike variants (SARS-CoV-2) Pseudotyped lentivirus (Luc reporter). It is a great tool to screen for variant-specific antibodies or to test the binding or efficacy of drug candidates against the different Spike variants. The Spike (SARS-CoV-2) pseudotyped lentiviruses can be used to measure the activity of neutralizing antibody against SARS-CoV-2 infection in a Biosafety Level 2 facility.
TCR/B2M Knockout NFAT Luciferase Reporter Jurkat Cell Line
Description: This cell line is a double knockout of TCR (T Cell Receptor) and B2M (Beta-2-Microglobulin). First, the TRAC (T-Cell Receptor Alpha Constant) and the TRBC1 (T-Cell Receptor Beta Constant 1) domains of the TCRα/β chains were genetically removed by CRISPR/Cas9 genome editing from NFAT Luciferase Reporter Jurkat cells to generate the TCR Knockout NFAT Luciferase Reporter Jurkat cell Line (BPS Bioscience #78556). These TCR knockout cells were used to generate a new cell line in which B2M was also genetically removed by CRISPR/Cas9 genome editing. _x000D_Expression of the firefly luciferase gene is driven by NFAT response elements located upstream of the minimal TATA promoter. Activation of the NFAT signaling pathway in these cells can be monitored by measuring luciferase activity.
Description: Recombinant Jurkat T cell expressing firefly luciferase gene under the control of NFAT response elements with constitutive expression of human CD160. CD160 is a GPIanchored glycoprotein member of the Ig superfamily, also known as BY55, NK1, and NK28. GenBank Accession # NM_007053._x000D__x000D_
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BQ.1 omicron variant has additional mutations (K444T and N460K) based on the BA.5 variant. The Spike (BQ.1, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BQ.1 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BQ.1, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BQ.1 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BQ.1 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BQ.1 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, K444T, L452R, N460K, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BQ.1 omicron variant has additional mutations (K444T and N460K) based on the BA.5 variant. The Spike (BQ.1, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BQ.1 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BQ.1, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BQ.1 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BQ.1 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BQ.1 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, K444T, L452R, N460K, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BQ.1.1 omicron variant has additional mutations (R346T, K444T and N460K) based on the BA.5 variant. The Spike (BQ.1.1, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BQ.1.1 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BQ.1.1, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BQ.1.1 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion_x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BQ.1.1 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BQ.1.1 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, R346T, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, K444T, L452R, N460K, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BQ.1.1 omicron variant has additional mutations (R346T, K444T and N460K) based on the BA.5 variant. The Spike (BQ.1.1, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BQ.1.1 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BQ.1.1, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BQ.1.1 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion_x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BQ.1.1 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BQ.1.1 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, R346T, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, K444T, L452R, N460K, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BF.7 omicron variant has additional mutation R346T based on the BA.5 variant. The Spike (BF.7, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BF.7 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BF.7, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BF.7 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BF.7 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BF.7 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, R346T, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BF.7 omicron variant has additional mutation R346T based on the BA.5 variant. The Spike (BF.7, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BF.7 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BF.7, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BF.7 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BF.7 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BF.7 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, R346T, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and human ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of January 2023, additional new sub-lineages (e.g. BQ.1, BQ.1.1, BF.7, XBB.1, XBB.1.5) have been designated._x000D_The Spike (XBB.1.5, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the XBB.1.5 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (XBB.1.5, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron XBB.1.5 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figure 2, the Spike Omicron XBB.1.5 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in XBB.1.5 Omicron Variant: T19I, LPP24-26del, A27S, V83A, G142D, Y144del, H146Q, Q183E, V213E, G252V, G339H, R346T, L368I, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, V445P, G446S, N460K, S477N, T478K, E484A, F486P, F490S, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and human ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of January 2023, additional new sub-lineages (e.g. BQ.1, BQ.1.1, BF.7, XBB.1, XBB.1.5) have been designated._x000D_The Spike (XBB.1.5, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the XBB.1.5 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (XBB.1.5, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron XBB.1.5 variant in a Biosafety Level 2 facility._x000D_
_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figure 2, the Spike Omicron XBB.1.5 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in XBB.1.5 Omicron Variant: T19I, LPP24-26del, A27S, V83A, G142D, Y144del, H146Q, Q183E, V213E, G252V, G339H, R346T, L368I, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, V445P, G446S, N460K, S477N, T478K, E484A, F486P, F490S, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K
Description: The Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) was produced with SARS-CoV-2 Spike corresponding to the initial strain (Genbank Accession #QHD43416.1) as the envelope glycoprotein instead of VSV-G. The pseudovirions contain the firefly luciferase gene; therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 in a Biosafety Level 2 facility.The Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) has been validated for use with target cells Vero-E6, Calu-3, and ACE2-HEK293 (BPS Bioscience #79951). Spike VSV Delta G are preferred for use in cells such as Vero-E6 and Calu-3.The infectivity of VSV-Delta G pseudotypes is restricted to a single round of replication, therefore the pseudotypes can be handled using BSL-2 containment practices.
Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter)
Description: The Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) was produced with SARS-CoV-2 Spike corresponding to the initial strain (Genbank Accession #QHD43416.1) as the envelope glycoprotein instead of VSV-G. The pseudovirions contain the firefly luciferase gene; therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 in a Biosafety Level 2 facility.The Spike (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) has been validated for use with target cells Vero-E6, Calu-3, and ACE2-HEK293 (BPS Bioscience #79951). Spike VSV Delta G are preferred for use in cells such as Vero-E6 and Calu-3.The infectivity of VSV-Delta G pseudotypes is restricted to a single round of replication, therefore the pseudotypes can be handled using BSL-2 containment practices.
Description: The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) was produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1; with D614G mutation) as the envelope glycoprotein instead of VSV-G. The pseudovirions contain the firefly luciferase gene; therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 D614G variant in a Biosafety Level 2 facility.The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) has been validated for use with target cells Vero-E6 and ACE2-HEK293 (BPS Bioscience #79951). Spike VSV Delta G is preferred over lentiviral-based spike pseudoviruses for use in cells such as Vero-E6 parental cells.
Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter)
Description: The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) was produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1; with D614G mutation) as the envelope glycoprotein instead of VSV-G. The pseudovirions contain the firefly luciferase gene; therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 D614G variant in a Biosafety Level 2 facility.The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) has been validated for use with target cells Vero-E6 and ACE2-HEK293 (BPS Bioscience #79951). Spike VSV Delta G is preferred over lentiviral-based spike pseudoviruses for use in cells such as Vero-E6 parental cells.
We show that these truncated GFPs may bind more non-native sequences, and this promiscuity invites the Possibility of a rational design of optimized sequences for fixing strands and photodissociation, both useful for optogenetic applications.
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