ACTC1 CRISPR Activation Plasmid (h): sc-400008-ACT

ACTC1 encodes alpha cardiac actin, a core component of the sarcomeric thin filament that polymerizes into F-actin to support force generation and contractile mechanics in striated muscle. By interacting with myosin motors and actin-binding regulators, ACTC1 contributes to myofibril assembly, cytoskeletal organization, and mechanotransduction pathways that coordinate cardiomyocyte structure and function. Perturbation of ACTC1 expression or filament integrity can alter sarcomere dynamics and calcium-handling coupling, linking this gene to inherited cardiomyopathy phenotypes and congenital cardiac defects. As a muscle-enriched actin isoform, ACTC1 is also used to study isoform-specific cytoskeletal remodeling and stress responses in cardiac and skeletal muscle models.

ACTC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACTC1 expression without altering the underlying DNA sequence.

ACTC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACTC1 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 ACTC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACTC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACTC1 locus and enabling the study of ACTC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACTC1 pathway restoration in tumor cells with silenced or reduced ACTC1 expression.

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