TCP-1 ζ CRISPR Activation Plasmid (h): sc-403047-ACT

CCT6A encodes the human chaperonin-containing TCP-1 subunit zeta (TCP-1 ζ), a core component of the cytosolic TRiC/CCT complex that mediates ATP-dependent folding of actin, tubulin, and additional client proteins required for proteostasis. Through its role in protein quality control, TCP-1 ζ supports cytoskeletal organization, cell-cycle progression, and stress-responsive homeostasis, with downstream effects on processes such as mitotic spindle integrity and intracellular trafficking. Perturbation of TRiC/CCT subunits can reshape signaling networks linked to proliferation and survival, and altered expression of CCT6A has been reported in multiple disease-relevant contexts, including cancer-associated phenotypes and proteostasis imbalance. As a result, CCT6A is frequently studied in pathways connecting chaperone capacity, cytoskeletal dynamics, and cellular fitness under stress.

TCP-1 ζ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCT6A expression without altering the underlying DNA sequence.

TCP-1 ζ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCT6A 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 CCT6A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCP-1 ζ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCT6A locus and enabling the study of TCP-1 ζ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCP-1 ζ pathway restoration in tumor cells with silenced or reduced CCT6A expression.

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