The purpose of this study was to investigate the expression level of miR-146a and miR-155 in cardiac xenograft models treated with immunosuppressant FK506, and to construct lentiviral vectors to further study the role of miR-146a and miR-155 in cardiac xenotransplantation. the expression levels of miR-146a and miR-155 is checked by quantitative polymerase chain reaction analysis and protein expression RELA, which is a member of the family of nuclear factor-kB, is checked by western blot analysis. Pre-miR-146a and miR-155 pre-designed and synthesized fragments according to MiRBase and cloned into plasmid pCDH1-MCS1-EF1-copGFP. recombinant plasmids were identified by enzyme digestion and sequencing.
Heart transplant survival time in FK506 treatment group increased significantly compared with the control group (P <0.05). In addition, the results of the histopathological grading was significantly decreased in the treatment group (P <0.05). A significant reduction in the level of protein expression Rela observed in the treatment group (P <0.05), along with a significant increase in the level of miR-146a expression (P <0.05) and a significant reduction in miR-155 expression levels ( P <0.05).
Digestion and sequencing findings suggest that miRNA insertion into plasmid pCDH1-MCS1-EF1-copGFP in accordance with pre-miRNAs, and concentrated into a lentiviral vector titer of 5 x 107 IFU / ml. These findings indicate that FK506 is able to inhibit the effect of rejection in cardiac xenotransplantation model of rat-to-rat. FK506 treatment alter the expression level of miR-146a and miR-155, which indicates that they may have an important role in regulating the immune response to the effects of rejection. miR-146a and miR-155 successfully constructed lentiviral vectors for further experiments both in vitro and in vivo.
Initial graft size and not the innate immune response limit survival of engrafted neural stem cells.
Formulation Nonionic Surfactant vesicles (NISV) Prepared by Microfluidics for Therapeutic Delivery of siRNA into cancer cells.
Disturb small RNA (siRNA) has a vast potential as a therapeutic agent for reversible silencing of target genes anything interesting. The clinical application of siRNA requires the use of safe and effective delivery system. In this study, we investigated the use of non-ionic surfactant vesicles (NISV) for siRNA delivery. Various types of formulations NISV microfluidic synthesized by mixing and then evaluated for cytotoxicity and physiochemical properties.
NISV ability to carry and transfect siRNA targeting the green fluorescent protein (GFP) to the A549 that stably express GFP (copGFP-A549) were evaluated. Flow cytometry and Western blotting were used to study the expression of GFP knockdown, and significant knockdown was observed as a result of the delivery of siRNA into cells by NISV. This happens especially when using Tween 85, which is capable of reaching more than 70% GFP knockdown. NISV thus shown to provide a promising and effective platform for the delivery of siRNA therapies.The application of stem cells in the treatment of various degenerative diseases are very promising.
PB-EF1-GreenPuro-H1-MCS shRNA cloning and expression vector
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: The ANPRA/Aequorin expression vector is designed to co-express human atrial natriuretic peptide receptor A (ANPRA, also called natriuretic peptide receptor A/guanylate cyclase A) and jellyfish (Aequorea victoria) Aequorin in mammalian cells.
Description: GFP-Rac1 Expression Vector Set contains 3 vectors: Rac wild type, T17N dominant negative mutant, and Q61L constitutively active mutant. Each vector also contains a GFP reporter sequence.
Description: GFP-RhoA Expression Vector Set contains 3 vectors: RhoA wild type, T19N dominant negative mutant, and Q63L constitutively active mutant. Each vector also contains a GFP reporter sequence.
Description: GFP-Cdc42 Expression Vector Set contains 3 vectors: Cdc42 wild type, T17N dominant negative mutant, and Q61L constitutively active mutant. Each vector also contains a GFP reporter sequence.
Description: Active Rac1 Expression Vector Set contains 3 vectors expressing different constitutively active mutants of Rac1: Q61L, Q61L/F37A, and Q61L/Y40C.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Active H-Ras Expression Vector Set contains 3 vectors expressing different constitutively active mutants of H-Ras: V12, V12S35, and V12C40.
pSIH1-H1-siLuc-copGFP Packaged Positive Transduction Control
Description: Clone your gene of interest into this AAV Expression Vector, then co-transfect along with AAV packaging vectors into a packaging host cell line such as 293AAV.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Baculovirus cassette vector pAc-l-CH3 for the expression of human, humanized or chimeric IgG(lambda) in insect cells and secretion of assembled antibodies into the supernatant.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Clone your gene of interest into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Description: Clone your gene of interest and a gene-specific promoter into this Lentiviral Expression Vector, then co-transfect along with lentiviral packaging vectors into a packaging cell line such as 293T or 293LTV. This expression vector is compatible with any 2nd or 3rd generation lentiviral packaging system, but due to its design it is best matched with our ViraSafe packaging vectors to produce the highest viral titer.
Excision only piggyBac Transposase expression vector, 50 rxns
Description: Human BRAF, also known as B-RAF1 and RAFB1, with 18 mutations to improve expression; S446A, S447A, I543A, I544S, I551K, Q562R, L588N, K630S, F667E, Y672S, A688R, L706S, Q709R, S713E, L716E, S720E, P722S, K723G. GenBank Accession #NM_004333, a.a. 444-723 with N-terminal His-tag, expressed in E. coli cells. MW = 33 kDa.
Description: Recombinant clonal CHO cell line stably expressing full-length human CEACAM5 (NM_004363). Surface expression of CEACAM5 was confirmed by flow cytometry. The stable clonal cell line was selected for different levels of CEACAM5 expression (High, Medium, and Low) compared to the parental CHO-K1 cell line.
Description: B7 homolog 3 protein (B7-H3) CHO Cell line is a recombinant clonal stable CHO cell line expressing full length human B7-H3 protein, also known as cluster of differentiation 276 (CD276, B7H3; B7RP-2; 4Ig-B7-H3). Surface expression of B7-H3 was confirmed by flow cytometry. Each stable clonal cell line was selected for High, Medium, or Low levels of B7-H3 expression to mimic different B7-H3 expression levels in various cancer types.
Description: Recombinant clonal HEK293 cell line stably expressing full length human CD36. The stable clonal cell line was selected for medium and high levels of CD36 expression compared to the parental HEK293 cell line.
Description: Recombinant clonal CHO cell line stably expressing full-length human CEACAM5 (NM_004363). Surface expression of CEACAM5 was confirmed by flow cytometry. The stable clonal cell line was selected for different levels of CEACAM5 expression (High, Medium, and Low) compared to the parental CHO-K1 cell line.
Description: The AAV Helper Free System produces recombinant AAV containing your gene of interest without the need to use a helper adenovirus. The adenoviral genes required for proper AAV packaging are provided in the pHelper plasmid (E2A, E4 and VA RNA) or in the 293 packaging cells (E1).
Description: The AAV Helper Free System produces recombinant AAV containing your gene of interest without the need to use a helper adenovirus. The adenoviral genes required for proper AAV packaging are provided in the pHelper plasmid (E2A, E4 and VA RNA) or in the 293 packaging cells (E1).
Description: The AAV Helper Free System produces recombinant AAV containing your gene of interest without the need to use a helper adenovirus. The adenoviral genes required for proper AAV packaging are provided in the pHelper plasmid (E2A, E4 and VA RNA) or in the 293 packaging cells (E1).
Description: The AAV Helper Free System produces recombinant AAV containing your gene of interest without the need to use a helper adenovirus. The adenoviral genes required for proper AAV packaging are provided in the pHelper plasmid (E2A, E4 and VA RNA) or in the 293 packaging cells (E1).
Description: The AAV Helper Free System produces recombinant AAV containing your gene of interest without the need to use a helper adenovirus. The adenoviral genes required for proper AAV packaging are provided in the pHelper plasmid (E2A, E4 and VA RNA) or in the 293 packaging cells (E1).
Description: The AAV Helper Free System produces recombinant AAV containing your gene of interest without the need to use a helper adenovirus. The adenoviral genes required for proper AAV packaging are provided in the pHelper plasmid (E2A, E4 and VA RNA) or in the 293 packaging cells (E1).
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However, the cell-based therapies can be limited by the problem of low survival of cells grafted and uncertainty about their fate. The combination of molecular imaging and contrast-enhanced MRI can provide more insight into the survival and behavior of transplanted stem cells. We explored the hair-follicle stem cell-derived bulge (HFBSCs) as a potential candidate for autologous cell-based therapies. HFBSCs were transduced with lentiviral construct to the gene coding for the bioluminescent (Luc2) and fluorescent (copGFP) reporter protein, and then loaded with magnetic nanoparticles to enable MRI visualization.
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