ChIP H3K36Me3 Mouse

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 Anti-acetyl-Histone H4 Antibody to perform ChIP acH4 - Rabbit Sheep YFP Tag

Get tips on using Anti-acetyl-Histone H3 (Lys9) Antibody to perform ChIP Anti-bodies H3K9-Ac

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

Get tips on using GATA-3 Antibody (HG3-31): sc-268 to perform ChIP Anti-bodies GATA3

Get tips on using HDAC1 (D5C6U) XP® Rabbit mAb #34589 to perform ChIP Anti-bodies HDAC1

Get tips on using SimpleChIP® Plus Sonication Chromatin IP Kit #56383 to perform ChIP Human - SW480

Get tips on using SimpleChIP® Plus Sonication Chromatin IP Kit #56383 to perform ChIP Human - AGS

Get tips on using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 to perform ChIP Human - ASM