siRNA / miRNA gene silencing Human PANC-1

Get tips on using siGENOME Human DAB2 (1601) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - A549 DAB2

Get tips on using Silencer® Select Negative Control No 1 siRNA to perform siRNA / miRNA gene silencing Mouse - siRNA negative control polymer / lipid

Get tips on using s15827 to perform siRNA / miRNA gene silencing Human - Capan-1 CDC7

Get tips on using SLC29A4 to perform siRNA / miRNA gene silencing Human - Capan-1 hENT1

Get tips on using ON-TARGETplus Human ARL2BP siRNA to perform siRNA / miRNA gene silencing Human - HeLa BART/ARL2BP

Get tips on using Silencer® GAPDH siRNA (human) to perform siRNA / miRNA gene silencing Human - Jurkat GAPDH 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 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 Human PCSK6 (5046) siRNA - Individual to perform siRNA / miRNA gene silencing Human - A253 PACE4

Get tips on using siGENOME Human FTO (79068) siRNA - Individual to perform siRNA / miRNA gene silencing Human - SHSY5Y FTO

Get tips on using siGENOME Human TBX2 (6909) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - U251 TBX2