siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line)

Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - PC-3 human prostate cancer

Get tips on using Human Breast Cancer Susceptibility Protein 2 (BRCA2) ELISA Kit to perform ELISA Human - BRCA2

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 YAP siRNA (h) to perform siRNA / miRNA gene silencing Human - OV2008 Yap Gene

Get tips on using siRNA P300 to perform siRNA / miRNA gene silencing Human - BOSC23 p300

Get tips on using MISSION® esiRNA_ human CCL2 to perform siRNA / miRNA gene silencing Human - U251 CCL2

Get tips on using siGENOME PCNT siRNA to perform siRNA / miRNA gene silencing Human - HEK293 PCNT

Get tips on using PCSK9 siRNA (h) to perform siRNA / miRNA gene silencing Human - A549 PCSK9

Get tips on using AmpFLSTR™ Identifiler™ Plus PCR Amplification Kit to perform Cell line authentication Human prostatic cancer cell line DU145