CARP CRISPR Activation Plasmid (h): sc-401842-ACT

Human ANKRD1 encodes cardiac ankyrin repeat protein (CARP), a stress-responsive transcriptional cofactor enriched in striated muscle that shuttles between the sarcomere and nucleus. CARP integrates mechanical and hypertrophic cues by modulating gene expression programs linked to sarcomere organization, cytoskeletal remodeling, and cardiac muscle development, including interactions with transcriptional regulators such as YAP/TEAD- and NF-κB-associated signaling. Altered ANKRD1/CARP expression has been associated with cardiomyopathy-related remodeling and broader muscle stress phenotypes, making it a useful target for dissecting mechanotransduction and transcriptional adaptation. In vitro models leveraging ANKRD1 perturbation are commonly applied to study contractile gene networks, myofibrillar integrity, and stress-induced transcriptomic shifts.

CARP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANKRD1 expression without altering the underlying DNA sequence.

CARP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANKRD1 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 ANKRD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CARP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANKRD1 locus and enabling the study of CARP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CARP pathway restoration in tumor cells with silenced or reduced ANKRD1 expression.

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