Calponin 2 CRISPR Activation Plasmid (h): sc-402882-ACT

Human CNN2 encodes calponin 2, an actin-binding regulatory protein that modulates cytoskeletal organization, actomyosin contractility, and cell shape. Calponin 2 associates with stress fibers and smooth muscle–like contractile structures, influencing processes such as cell adhesion, migration, and mechanotransduction through actin–myosin dynamics and Rho GTPase–linked signaling. By tuning cytoskeletal tension and motility programs, CNN2 has been studied in contexts including vascular and inflammatory cell behavior, fibrosis-associated remodeling, and tumor cell invasion and metastasis. Altered calponin 2 expression or localization can therefore serve as a mechanistic readout for changes in cytoskeletal pathways that impact tissue remodeling and disease-related cellular phenotypes.

Calponin 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CNN2 expression without altering the underlying DNA sequence.

Calponin 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CNN2 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 CNN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Calponin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CNN2 locus and enabling the study of Calponin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Calponin 2 pathway restoration in tumor cells with silenced or reduced CNN2 expression.

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