p41-ARCb CRISPR Activation Plasmid (h): sc-405027-ACT

ARPC1B encodes p41-ARCb, a regulatory subunit of the Arp2/3 complex that promotes branched actin polymerization required for lamellipodia formation, endocytosis, and dynamic remodeling of the cortical cytoskeleton. Through coupling actin nucleation to signaling inputs, ARPC1B supports processes such as cell migration, immune synapse organization, and vesicular trafficking. Altered ARPC1B function is associated with dysregulated actin-dependent responses in hematopoietic and immune cell contexts, making it relevant for studying cytoskeletal control of inflammation and host defense. ARPC1B is therefore frequently investigated in pathways governing cell shape changes, adhesion, and receptor-driven signaling that depend on Arp2/3 activity.

p41-ARCb CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARPC1B expression without altering the underlying DNA sequence.

p41-ARCb CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARPC1B 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 ARPC1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p41-ARCb expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARPC1B locus and enabling the study of p41-ARCb-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p41-ARCb pathway restoration in tumor cells with silenced or reduced ARPC1B expression.

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