Arp3β CRISPR Activation Plasmid (h): sc-402186-ACT

ACTR3B encodes Arp3β, an actin-related protein that is thought to contribute to dynamic remodeling of the actin cytoskeleton, supporting processes such as cell shape control, membrane ruffling, endocytosis, and cell migration. As part of actin regulatory networks linked to Arp2/3-dependent nucleation and branched actin assembly, Arp3β can influence cortical actin organization and cytoskeletal responses to signaling cues. Perturbation of actin polymerization pathways is broadly relevant to mechanisms underlying invasive behavior, altered adhesion, and dysregulated intracellular trafficking observed in multiple disease contexts. ACTR3B expression and functional modulation are therefore useful entry points for studying cytoskeleton-centric phenotypes and pathway connectivity in human cell models.

Arp3β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACTR3B expression without altering the underlying DNA sequence.

Arp3β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACTR3B 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 ACTR3B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Arp3β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACTR3B locus and enabling the study of Arp3β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Arp3β pathway restoration in tumor cells with silenced or reduced ACTR3B expression.

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