Calponin 1 CRISPR Activation Plasmid (m): sc-419719-ACT

Mouse Cnn1 encodes calponin 1, an actin-binding protein enriched in differentiated smooth muscle where it modulates contractile behavior by regulating actin–myosin interactions and cytoskeletal organization. Calponin 1 participates in calcium-dependent control of actomyosin dynamics and is linked to RhoA/ROCK signaling, focal adhesion remodeling, and phenotypic switching between contractile and synthetic smooth muscle states. Changes in CNN1 expression are used as readouts of smooth muscle differentiation and vascular remodeling, with relevance to processes such as neointimal hyperplasia, fibrosis-associated myofibroblast programs, and alterations in tissue biomechanics. As a cytoskeletal regulator, calponin 1 provides a functional node for studying mechanotransduction, migration, and extracellular matrix–driven signaling in vascular and airway smooth muscle models.

Calponin 1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cnn1 expression without altering the underlying DNA sequence.

Calponin 1 CRISPR Activation Plasmid (m) 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.