
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IRS-2 CRISPR Activation Plasmid (h) | sc-400474-ACT | 20 µg | $397.00 |
Human IRS2 encodes insulin receptor substrate 2 (IRS-2), a cytosolic adaptor that becomes tyrosine-phosphorylated downstream of insulin and IGF1 receptors to couple receptor signaling to PI3K–AKT and MAPK cascades. Through recruitment of SH2-domain effectors, IRS-2 integrates cues that control glucose metabolism, cell growth, survival, and differentiation, with prominent roles in hepatic insulin action and β-cell function. Altered IRS2 signaling is frequently studied in the context of insulin resistance, obesity-associated metabolic dysfunction, and signaling rewiring in cancer biology. IRS-2 also interfaces with nutrient-sensing and inflammatory inputs that modulate transcriptional programs and cellular stress responses.
IRS-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IRS2 expression without altering the underlying DNA sequence.
IRS-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IRS2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the IRS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IRS-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IRS2 locus and enabling the study of IRS-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IRS-2 pathway restoration in tumor cells with silenced or reduced IRS2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.