ChIP acH3 Rat Sheep BEF Tag

Get tips on using CtIP antibody (mAb) to perform ChIP Anti-bodies CtIP/BCL11A

Get tips on using MAGnify™ Chromatin Immunoprecipitation System to perform ChIP Mouse - RAW264.7

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Mouse - RAW264.7

Get tips on using Bcl-11B (D6F1) XP® Rabbit mAb #12120 to perform ChIP Anti-bodies CtIP/BCL11A

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 Mre11 Antibody #4895 to perform ChIP Anti-bodies MRE11

Get tips on using Anti-CTCF Antibody to perform ChIP Anti-bodies CTCF

Get tips on using ChromaFlash Chromatin Extraction Kit to perform ChIP Mouse - Osteoblasts

Get tips on using CTCF (D31H2) XP® Rabbit mAb #3418 to perform ChIP Anti-bodies CTCF

Get tips on using Estrogen Receptor α (D8H8) Rabbit mAb #8644 to perform ChIP Anti-bodies ERα