ACTG2 CRISPR Activation Plasmid (h): sc-400007-ACT

ACTG2 encodes gamma-2 smooth muscle actin, a core component of the contractile apparatus in visceral smooth muscle cells where it polymerizes into actin filaments that support force generation and cytoskeletal integrity. ACTG2 participates in actomyosin dynamics and cytoskeleton-dependent processes including smooth muscle contraction, cell shape control, and mechanotransduction, interacting functionally with myosin and actin-binding proteins across RhoA/ROCK-regulated pathways. Altered ACTG2 expression or function is associated with impaired gastrointestinal and genitourinary smooth muscle motility phenotypes and has been implicated in congenital visceral myopathy spectrum disorders. As a lineage- and state-associated smooth muscle marker, ACTG2 is also used to study differentiation programs, cytoskeletal remodeling, and contractility-driven cellular phenotypes.

ACTG2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACTG2 expression without altering the underlying DNA sequence.

ACTG2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACTG2 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 ACTG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACTG2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACTG2 locus and enabling the study of ACTG2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACTG2 pathway restoration in tumor cells with silenced or reduced ACTG2 expression.

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