siRNA / miRNA gene silencing Human OV2008 Yap Gene

Get tips on using Brn-3b siRNA (m) to perform siRNA / miRNA gene silencing Rat - Retinal stem cells Brn-3b

Get tips on using HIF-1α siRNA (r) to perform siRNA / miRNA gene silencing Rat - Cardiomyocyte (H9C2) HIF-1α Lipid

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 have 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 efficacy of transduction and shRNA on its target site.

Get tips on using FlexiTube GeneSolution GS26354 for GNL3 to perform siRNA / miRNA gene silencing Human - MDA-MB-231 GNL3

Get tips on using ON-TARGETplus Mouse Jun siRNA to perform siRNA / miRNA gene silencing Mouse - Embryonic stem cells Jun

Get tips on using ON-TARGETplus Mouse Gpam siRNA to perform siRNA / miRNA gene silencing Mouse - Embryonic stem cells Gpat1

Get tips on using Silencer® Select_Mapk14/p38 siRNA(r) to perform siRNA / miRNA gene silencing Rat - NRVM( Mapk14/p38

Get tips on using siGENOME Rat Arhgap35 (306400) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 p190RhoGAP/Arhgap35

Get tips on using siGENOME Rat Sod2 (24787) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - NRK MnSOD/Sod2

Get tips on using siGENOME Mouse Sod2 (20656) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RGC-5 Sod2