siRNA / miRNA gene silencing Human TF‐1

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.

Get tips on using ON-TARGETplus Rat Arhgef6 (363509) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 α-Pix/Arhgef6

Get tips on using ON-TARGETplus Rat Itgb1 (24511) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 β1 integrin/Itgb1

Get tips on using ON-TARGETplus Rat Tp53 (24842) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - C6 (rat glioma) p53

Get tips on using ON-TARGETplus Mouse Mapk1 (26413) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 MAPK1 (ERK2)

Get tips on using ON-TARGETplus Mouse Mapk14 (26416) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 p38/Mapk14

Get tips on using ON-TARGETplus Mouse Nr1h3 (22259) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - M210B4 LXR‐α/Nr1h3

Get tips on using ON-TARGETplus Mouse Nr1h2 (22260) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - M210B4 LXR‐β/Nr1h2

Get tips on using ON-TARGETplus Mouse Zbp1 (58203) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 3T3-SA Zbp1/Dai

Get tips on using Silencer® Select_FPr1/Silencer® Select_FPr2 siRNA to perform siRNA / miRNA gene silencing Mouse - Neuro 2a Fpr1/Fpr2