Arp2 CRISPR Activation Plasmid (h): sc-401018-ACT

ACTR2 encodes the human actin-related protein Arp2, a core subunit of the Arp2/3 complex that nucleates branched actin filaments to remodel the cytoskeleton. Arp2/3-dependent actin dynamics drive lamellipodia formation, cell migration, endocytosis, and phagocytosis, and they coordinate with Rho-family GTPase signaling and adhesion pathways to regulate morphogenesis and intracellular trafficking. Through its role in shaping actin networks, ACTR2 influences processes such as immune cell motility, epithelial barrier remodeling, and vesicle transport, making it relevant to studies of invasion, metastasis-associated phenotypes, and neurodevelopmental or cytoskeletal dysfunction. Perturbations in actin assembly pathways are frequently observed in cancers and inflammatory contexts, and ACTR2 provides a mechanistic entry point for dissecting these cytoskeleton-linked changes.

Arp2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACTR2 expression without altering the underlying DNA sequence.

Arp2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACTR2 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 ACTR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Arp2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACTR2 locus and enabling the study of Arp2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Arp2 pathway restoration in tumor cells with silenced or reduced ACTR2 expression.

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