IGF2R/M6PR CRISPR Activation Plasmid (h): sc-401697-ACT

IGF2R encodes the cation-independent mannose-6-phosphate receptor (IGF2R/M6PR), a multifunctional endosomal sorting receptor that traffics M6P-tagged lysosomal hydrolases from the Golgi to endosomes and lysosomes while also mediating uptake and clearance of extracellular ligands. By binding insulin-like growth factor 2 (IGF2) and regulating ligand availability, IGF2R influences growth factor signaling networks and cellular proliferation programs. IGF2R/M6PR participates in lysosome biogenesis, endosome-to-Golgi recycling, and proteostasis pathways that shape metabolism and stress responses. Dysregulation or loss of IGF2R function has been associated with altered lysosomal enzyme trafficking and aberrant IGF2 signaling, contexts frequently explored in cancer biology, developmental biology, and lysosomal dysfunction research.

IGF2R/M6PR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IGF2R expression without altering the underlying DNA sequence.

IGF2R/M6PR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IGF2R 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 IGF2R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IGF2R/M6PR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IGF2R locus and enabling the study of IGF2R/M6PR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IGF2R/M6PR pathway restoration in tumor cells with silenced or reduced IGF2R expression.

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