siRNA / miRNA gene silencing Human siRNA negative control

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1) Lipid

Get tips on using siRNA Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - IEC Cationic lipid based

Get tips on using siRNA Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - IEC-6 Cationic lipid based

Get tips on using MISSION® esiRNA_esiRNA targeting mouse Lrp5 (esiRNA1) to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Lrp5

Get tips on using MISSION® esiRNA_esiRNA targeting mouse Lrp6 (esiRNA1) to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Lrp6

Get tips on using pSUPER.retro.neo+gfp vector- Syn G (exon 3) siRNA to perform shRNA gene silencing Mouse - RGC-5 Syn G (Exon 3)

Get tips on using TaqMan® MicroRNA Reverse Transcription Kit to perform siRNA / miRNA gene silencing Mouse - Glomerular mesangial cells HIPK2 Polymer / Lipid delivery

Get tips on using X-tremeGENE™ siRNA Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - AR42J Lipid based

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.

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.