IGFBP5 CRISPR Activation Plasmid (h): sc-400945-ACT

IGFBP5 (insulin-like growth factor binding protein 5) is a secreted IGF-binding protein that modulates IGF1/IGF2 bioavailability and signaling through the IGF axis, influencing PI3K–AKT and MAPK pathway activity. By regulating growth factor responsiveness and extracellular matrix interactions, IGFBP5 contributes to control of cell proliferation, survival, differentiation, migration, and tissue remodeling. Altered IGFBP5 expression has been linked to changes in fibrosis-associated programs, stromal biology, and tumor microenvironment dynamics, making it a useful node for studying context-dependent growth regulation. As a human protein with both IGF-dependent and IGF-independent functions, IGFBP5 is frequently investigated in pathways governing cellular senescence and developmental or injury responses.

IGFBP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IGFBP5 expression without altering the underlying DNA sequence.

IGFBP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IGFBP5 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 IGFBP5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IGFBP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IGFBP5 locus and enabling the study of IGFBP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IGFBP5 pathway restoration in tumor cells with silenced or reduced IGFBP5 expression.

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