IGFBP6 CRISPR Activation Plasmid (h): sc-404768-ACT

IGFBP6 encodes insulin-like growth factor binding protein 6, a secreted glycoprotein that preferentially binds IGF-II and modulates its bioavailability, receptor engagement, and downstream signaling through pathways such as PI3K–AKT and MAPK/ERK. By regulating IGF-dependent growth, survival, and differentiation cues, IGFBP6 contributes to tissue homeostasis and cellular stress responses in multiple organ contexts. Altered IGFBP6 expression has been reported across diverse cancer and metabolic research settings, where it can influence proliferation, migration, and apoptosis phenotypes linked to dysregulated IGF signaling. As a soluble extracellular regulator, IGFBP6 is also studied for its impact on paracrine communication within the tumor microenvironment and in developmental and endocrine biology models.

IGFBP6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IGFBP6 expression without altering the underlying DNA sequence.

IGFBP6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IGFBP6 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 IGFBP6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IGFBP6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IGFBP6 locus and enabling the study of IGFBP6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IGFBP6 pathway restoration in tumor cells with silenced or reduced IGFBP6 expression.

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