ChIP H3K27me3 Human Goat

Get tips on using ON-TARGETplus Human RAB11FIP1 (80223) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - A431 Rab Coupling Protein (RCP)

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 Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb #5326 to perform ChIP Anti-bodies H3K4me1

Get tips on using ON-TARGETplus Human ITGB3 (3690) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MDA-MB-231 β3 integrin/ITGB3

Get tips on using ON-TARGETplus Human ITGB1 (3688) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MDA-MB-231 β1 integrin/ITGB1

Get tips on using ON-TARGETplus Human LIN28A (79727) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - hES cell line H1 (WA01) LIN28

Get tips on using StemMACS™ AdipoDiff Media, human to perform Stem cell Differentiation media hMSCs differentiation into adipogenic cells