p21-ARC CRISPR Activation Plasmid (h): sc-403677-ACT

ARPC3 encodes p21-ARC, an essential subunit of the Arp2/3 complex that nucleates branched actin filament networks to drive lamellipodia formation, membrane protrusion, and force generation during cell migration. Through coordinated regulation by Rho-family GTPases and nucleation-promoting factors such as the WAVE and WASP families, p21-ARC supports actin-dependent processes including endocytosis, phagocytosis, vesicle trafficking, and immunological synapse organization. Perturbation of Arp2/3-mediated cytoskeletal remodeling influences epithelial morphogenesis, neuronal outgrowth, and leukocyte motility, linking ARPC3 activity to mechanisms commonly leveraged in invasion and metastasis biology. As a core actin regulator, ARPC3 is frequently studied in pathways controlling cell shape, adhesion turnover, and mechanotransduction under normal and disease-relevant conditions.

p21-ARC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARPC3 expression without altering the underlying DNA sequence.

p21-ARC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARPC3 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 ARPC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p21-ARC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARPC3 locus and enabling the study of p21-ARC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p21-ARC pathway restoration in tumor cells with silenced or reduced ARPC3 expression.

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