ELISA (kit) Human Serum Cytokine measurements (Multiplex assay) -NA- Human

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-Type I Collagen to perform Immunohistochemistry Collagen Type I - Goat Mouse -NA-

Get tips on using PDGF Receptor β (28E1) Rabbit mAb to perform Immunohistochemistry PDGFβR - Rabbit Mouse -NA-

Get tips on using Anti-trimethyl-Histone H3 (Lys9) Antibody to perform ChIP H3K9me3 - Sheep Rat -NA-

Get tips on using Anti-trimethyl-Histone H3 (Lys4) Antibody to perform ChIP H3K4Me3 - Sheep Rat -NA-

Get tips on using RNAprep pure Kit to perform RNA isolation / purification Tissue - Human Placenta

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - Human Lung

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Tissue - Human Liver

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - Human Liver