Troponin I-C CRISPR Activation Plasmid (h): sc-400466-ACT

TNNI3 encodes cardiac troponin I, the inhibitory subunit of the troponin complex that couples cytosolic Ca²⁺ dynamics to sarcomere activation in striated muscle. By modulating actin–myosin interaction through Ca²⁺-dependent regulation of tropomyosin positioning, Troponin I-C helps set contractile force and relaxation kinetics within the cardiomyocyte thin filament. TNNI3 function is integrated with excitation–contraction coupling, sarcomeric assembly, and phosphorylation-dependent signaling that tunes myofilament Ca²⁺ sensitivity. Genetic and regulatory perturbations of TNNI3 are linked to cardiomyopathy-related phenotypes and arrhythmogenic mechanisms, making it a central target for studying cardiac contractility and myofibrillar stress responses.

Troponin I-C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNNI3 expression without altering the underlying DNA sequence.

Troponin I-C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNNI3 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 TNNI3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Troponin I-C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNNI3 locus and enabling the study of Troponin I-C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Troponin I-C pathway restoration in tumor cells with silenced or reduced TNNI3 expression.

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