siRNA / miRNA gene silencing

Get tips on using β-Galactosidase Reporter Gene Staining Kit to perform Reporter gene assay β-galactosidase substrates - mouse embryo tissue

Get tips on using MISSION® shRNA SOX2 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans SOX2 lentiviral particles

Get tips on using MISSION® shRNA SOX6 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans SOX6 lentiviral particles

Get tips on using MISSION® shRNA ZEB2 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans ZEB2 lentiviral particles

Get tips on using MISSION® shRNA ZEB1 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans ZEB1 lentiviral particles

Get tips on using β-Gal Reporter Gene Assay, chemiluminescent to perform Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem cells

Get tips on using β-Galactosidase Reporter Gene Staining Kit to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells

Get tips on using β-Galactosidase Reporter Gene Staining Kit to perform Reporter gene assay β-galactosidase substrates - rat mesenchymal stem cells (MSCs)

Reporter gene assays are designed to test the regulation of the expression of a gene of interest. This is usually done by linking the promoter of the gene of interest with a gene such as a firefly luciferase, which can be easily detected by addition of luciferin that leads to an enzymatic reaction to produce luminescence. The enzymatic reaction can be correlated to the expression of the gene of interest. Another luciferase gene that can be used is Renilla luciferase. For an appropriate luciferase assay: 1. the reporter should express uniformly in all cells, 2. specifically respond to effectors that the assay intends to monitor, 3. have low intrinsic stability to quickly reflect transcriptional dynamics. It is important to have an equal number of cells plated in each testing condition to avoid any incorrect readouts. Reporter assays could be single or dual reporter assays. The reporter could be both luciferases. Most dual-luciferase assays involve adding two reagents to each sample and measuring luminescence following each addition. Adding the first reagent activates the first luciferase reporter reaction; adding the second reagent extinguishes first luciferase reporter activity and initiates the second luciferase reaction. Dual-luciferase assays have some advantages, including 1. reduces variability, 2. reduces background, 3. normalizes differences in transfection efficiencies between samples.

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase