Get tips on using ON-TARGET plus Mouse Becn1 (56208) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 4T1 BECN-1
Get tips on using SignalSilence® NF-κB p65 siRNA I #6261 to perform siRNA / miRNA gene silencing Rat - H9c2 NF-κB RelA (p65)
Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 ROCK2
Get tips on using pcDNA™3.1 (+) Mammalian Expression Vector to perform siRNA / miRNA gene silencing Human - U251 cofilin-1 (CFL1)
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 ON-TARGETplus Rat Snap23 (64630) siRNA - Set of 4 to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Snap23
Get tips on using heat shock protein family A (Hsp70) member 5 to perform siRNA / miRNA gene silencing Human - PC3 (human prostate cancer cell line) HSPA5 (GRP78)
Hello Iam a phd student in pharmacy and i want to know if this technology is suitable to knockout or silencing part of the gas5 gene in BV2 cells please
Get tips on using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb to perform siRNA / miRNA gene silencing Human - COV-434 SAPK/JNK
Get tips on using - to perform siRNA / miRNA gene silencing Rat - IEC-6
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