siRNA / miRNA gene silencing Rat Brain endothelial cells HIF-1α

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Get tips on using Endothelial Cell Growth Medium MV to perform Mammalian cell culture media BAOEC

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Get tips on using SASI_Hs01_00024301 to perform siRNA / miRNA gene silencing Human - MOLT4 RAG1

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Get tips on using Fenozol to perform siRNA / miRNA gene silencing Human - BOSC23

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Short hairpin or small hairpin RNA (shRNA) is artificial RNA, which has a hairpin loop structure, and uses inherent microRNA (miRNA) machinery to silence target gene expression. This is called RNA interference (RNAi). These can be delivered via plasmids or viral/bacterial vectors. Challenges in shRNA-mediated gene silencing include 1. Off-target silencing, 2. Packaging shRNA encoding lentivirus, and 3. Stable transduction in cells. RNAi has been designed to have anywhere from 19-27 bs, but the most effective design has 19 bp. In case commercial shRNAs are not available, potential target sites can be chosen within exon, 5’- or 3’ UTR, depending on which splice variants of the gene are desired. One should use the latest algorithms and choose at least two different sequences, targeting different regions, in order to have confidence in overcoming off-target effects. A BLAST search after selecting potential design will eliminate potential off-target sequences. For the second challenge, sequencing the vector using primers for either strand (50-100 bp upstream) is suggested, along with using enzymatic digestion on agarose gel for the vector. Next, once the shRNA-containing vector is packaged in a virus, it is important to check the viral titer before transduction. Finally, using a marker in the lentiviral vector (fluorescent protein or antibiotic resistance), along with qPCR for target gene expression can help in determining the efficacy of transduction and shRNA on its target site.

RNA shRNA gene silencing Human TF‐1 GATA‐1

Hello Iam a phd student in pharmacy and i want to know if this technology is suitable to knockout or silencing part of the gas5 gene in BV2 cells please

Discussions Bv2 GAS5 gene silencing

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human aortic endothelial cells

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Get tips on using Stealth siRNA_GATA2 to perform siRNA / miRNA gene silencing Human - LAD2 GATA2

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Get tips on using Neogen-Brain-Heart Infusion Broth to perform Bacterial cell culture media Salmonella enterica

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Get tips on using BHI-BRAIN HEART INFUSION BROTH to perform Bacterial cell culture media Listeria monocytogens

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