INSR/Insulin Receptor CRISPR Activation Plasmid (h): sc-400075-ACT

INSR encodes the insulin receptor, a transmembrane receptor tyrosine kinase that binds insulin to initiate autophosphorylation and downstream signaling through PI3K–AKT and RAS–MAPK pathways. These cascades coordinate glucose uptake, glycogen synthesis, lipid metabolism, and broader control of cell growth and survival, with cross-talk to mTOR and FOXO-regulated transcriptional programs. INSR activity shapes receptor endocytosis and recycling dynamics and influences insulin sensitivity at the level of adaptor proteins such as IRS family members. Dysregulation of INSR signaling is strongly associated with insulin resistance and metabolic disease biology and is also relevant to altered growth signaling observed in multiple tissues and tumor microenvironments.

INSR/Insulin Receptor CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous INSR expression without altering the underlying DNA sequence.

INSR/Insulin Receptor CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the INSR 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 INSR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous INSR/Insulin Receptor expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native INSR locus and enabling the study of INSR/Insulin Receptor-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of INSR/Insulin Receptor pathway restoration in tumor cells with silenced or reduced INSR expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.