siRNA / miRNA gene silencing Human PANC-1

Get tips on using p21 siRNA (h) to perform siRNA / miRNA gene silencing Human - COV-434 P21

Get tips on using Hs_HDAC2_1 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - LOR-L23 HDAC2_1

Get tips on using Hs_HDAC1_6 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - LOR-L23 HDAC1_6

Get tips on using Hs_BAP1_5 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - LOR-L23 BAP1_5

Get tips on using Hs_BAP1_3 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - LOR-L23 BAP1_3

Get tips on using siRNA TRAF6 (h) to perform siRNA / miRNA gene silencing Human - Calu-3 TRAF6

Get tips on using Nox3 siRNA (h) to perform siRNA / miRNA gene silencing Human - BEAS-2B NOX3

Get tips on using Nrf2 siRNA (m) to perform siRNA / miRNA gene silencing Human - BEAS-2B Nrf

Get tips on using siRNA p62 / SQSTM1 to perform siRNA / miRNA gene silencing Human - BEAS-2B P62

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.