CapZ-α1 CRISPR Activation Plasmid (h): sc-405348-ACT

CAPZA1 encodes CapZ-α1, an actin filament barbed-end capping protein that partners with β subunits to regulate actin polymerization dynamics, filament stability, and cytoskeletal remodeling. By controlling actin assembly at the leading edge and within stress fibers, CapZ-α1 influences cell shape, adhesion, and motility, integrating with pathways governing focal adhesion turnover and mechanotransduction. CAPZA1 activity is also linked to sarcomeric and contractile actin organization, supporting studies of cytoskeletal architecture in differentiated and proliferative cell states. Dysregulated actin capping and remodeling are frequently implicated in phenotypes relevant to oncogenic invasion, cardiomyocyte structural integrity, and other disorders where altered cytoskeletal mechanics contribute to pathology.

CapZ-α1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAPZA1 expression without altering the underlying DNA sequence.

CapZ-α1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAPZA1 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 CAPZA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CapZ-α1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAPZA1 locus and enabling the study of CapZ-α1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CapZ-α1 pathway restoration in tumor cells with silenced or reduced CAPZA1 expression.

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