SgK269 CRISPR Activation Plasmid (h): sc-403322-ACT

PEAK1 (also known as SgK269) encodes a cytoskeleton-associated pseudokinase that functions as a scaffold integrating signals from receptor tyrosine kinases and integrin-mediated adhesion. It regulates focal adhesion dynamics, actin remodeling, and cell motility through interactions that influence SRC/FAK signaling and downstream pathways such as MAPK/ERK and PI3K/AKT. PEAK1 contributes to invasive phenotypes by coordinating phosphorylation-dependent signaling complexes and modulating epithelial–mesenchymal transition-related processes. Altered PEAK1 expression and pathway engagement have been associated with tumor progression and metastatic behavior in multiple cancer contexts, supporting its use in mechanistic studies of adhesion and migration.

SgK269 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PEAK1 expression without altering the underlying DNA sequence.

SgK269 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PEAK1 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 PEAK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SgK269 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PEAK1 locus and enabling the study of SgK269-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SgK269 pathway restoration in tumor cells with silenced or reduced PEAK1 expression.

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