Telethonin CRISPR Activation Plasmid (h): sc-402317-ACT

TCAP encodes telethonin, a sarcomeric Z-disc protein that binds the N-terminus of titin (TTN) and helps stabilize myofibril assembly and mechanical coupling in striated muscle. Telethonin participates in sarcomere integrity and mechanotransduction-associated signaling, coordinating protein complex organization at the Z-disc during muscle development and adaptation. Perturbation of TCAP expression or telethonin function has been linked to inherited cardiomyopathies and skeletal muscle disorders, reflecting sensitivity of contractile architecture to Z-disc protein stoichiometry. As a node in titin-centered assemblies, telethonin is frequently studied in pathways governing cytoskeletal organization, muscle fiber maintenance, and stress-responsive remodeling.

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

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

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