TCP-1 η CRISPR Activation Plasmid (h): sc-403833-ACT

CCT7 encodes the TCP-1 η (CCTη) subunit of the cytosolic chaperonin containing TCP-1 (CCT/TRiC) complex, an ATP-dependent folding machine required for maturation of structurally complex proteins such as actin and tubulin. By supporting cytoskeletal assembly, proteostasis, and quality control, CCT/TRiC influences cell cycle progression, intracellular trafficking, and stress responses, including crosstalk with ubiquitin–proteasome pathways. Dysregulated chaperonin activity has been linked to altered protein homeostasis programs observed in proliferative and neurodegenerative disease contexts, making CCT7 a useful node for interrogating folding-dependent phenotypes. In human cells, perturbing CCT7 expression can reveal how chaperone capacity constrains cytoskeletal dynamics and signaling networks that depend on properly folded client proteins.

TCP-1 η CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCT7 expression without altering the underlying DNA sequence.

TCP-1 η CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCT7 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 CCT7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCP-1 η expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCT7 locus and enabling the study of TCP-1 η-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCP-1 η pathway restoration in tumor cells with silenced or reduced CCT7 expression.

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