p16-ARC CRISPR Activation Plasmid (h): sc-403469-ACT

Human ARPC5 encodes p16-ARC, a core subunit of the Arp2/3 complex that nucleates branched actin filaments to control lamellipodia formation, cell polarity, endocytosis, and vesicular trafficking. Through regulation of actin remodeling downstream of Rho-family GTPases, Arp2/3 activity coordinates adhesion turnover and directional migration, shaping cytoskeletal dynamics during proliferation and differentiation. Altered Arp2/3 complex function and actin network organization have been linked to invasive phenotypes and metastatic behavior across multiple cancer contexts, and to broader defects in membrane trafficking and immune cell motility. ARPC5 is therefore a useful node for studying cytoskeleton-dependent signaling, mechanotransduction, and microenvironment-driven changes in cell movement.

p16-ARC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARPC5 expression without altering the underlying DNA sequence.

p16-ARC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARPC5 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 ARPC5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p16-ARC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARPC5 locus and enabling the study of p16-ARC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p16-ARC pathway restoration in tumor cells with silenced or reduced ARPC5 expression.

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