Troponin C CRISPR Activation Plasmid (h): sc-406429-ACT

TNNC1 encodes cardiac/slow skeletal troponin C, a Ca2+-binding EF-hand protein that functions as the calcium sensor within the thin filament troponin complex to regulate actin–myosin interaction and sarcomere contraction. Upon Ca2+ binding, Troponin C drives conformational changes that relieve tropomyosin-mediated inhibition, coupling excitation–contraction signaling to force generation in striated muscle. TNNC1 activity intersects with calcium homeostasis and contractile apparatus pathways that shape myofibril assembly, muscle energetics, and mechanotransduction. Dysregulated TNNC1 expression or function is implicated in inherited cardiomyopathies and contractile phenotypes, supporting its use in studies of sarcomere biology and cardiac disease mechanisms.

Troponin C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNNC1 expression without altering the underlying DNA sequence.

Troponin C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNNC1 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 TNNC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Troponin C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNNC1 locus and enabling the study of Troponin C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Troponin C pathway restoration in tumor cells with silenced or reduced TNNC1 expression.

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