IMP-1/IGF2BP1 CRISPR Activation Plasmid (h): sc-401703-ACT

IGF2BP1 (IMP-1) is an RNA-binding protein that recognizes N6-methyladenosine (m6A) and other sequence/structure features to regulate mRNA localization, stability, and translation during development and cellular stress responses. By modulating the fate of transcripts linked to growth and metabolism, IGF2BP1 influences programs controlling proliferation, migration, and epithelial–mesenchymal transition, interfacing with signaling networks such as PI3K–AKT–mTOR and MAPK through downstream target expression. Aberrant IGF2BP1 expression has been reported in multiple tumor contexts and is associated with altered post-transcriptional control of oncogenic and pro-survival mRNAs. In addition, IGF2BP1-dependent RNA regulation is relevant to stem-like phenotypes and remodeling of cellular state, supporting mechanistic studies of gene regulatory networks.

IMP-1/IGF2BP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IGF2BP1 expression without altering the underlying DNA sequence.

IMP-1/IGF2BP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IGF2BP1 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 IGF2BP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IMP-1/IGF2BP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IGF2BP1 locus and enabling the study of IMP-1/IGF2BP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IMP-1/IGF2BP1 pathway restoration in tumor cells with silenced or reduced IGF2BP1 expression.

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