Calponin 1 CRISPR Activation Plasmid (h): sc-401397-ACT

CNN1 encodes calponin 1, an actin-binding regulatory protein enriched in smooth muscle and myofibroblast-like states where it modulates actomyosin contractility. By binding filamentous actin and influencing myosin ATPase activity, calponin 1 contributes to cytoskeletal organization, cell adhesion, and contractile force generation within pathways that coordinate RhoA/ROCK signaling and actin stress fiber dynamics. Altered CNN1 expression is used as a marker of smooth muscle differentiation and phenotypic switching, processes relevant to vascular remodeling and fibroproliferative responses. Dysregulation of calponin 1–associated cytoskeletal programs has been reported in contexts of abnormal tissue stiffness and motility, supporting its use in studies of cellular plasticity and mechanobiology.

Calponin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CNN1 expression without altering the underlying DNA sequence.

Calponin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CNN1 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 CNN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Calponin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CNN1 locus and enabling the study of Calponin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Calponin 1 pathway restoration in tumor cells with silenced or reduced CNN1 expression.

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