TCP-1 γ CRISPR Activation Plasmid (h): sc-403203-ACT

CCT3 encodes the human TCP-1 gamma subunit of the cytosolic chaperonin-containing TCP1 (CCT/TRiC) complex, an essential ATP-dependent folding machine for many newly synthesized proteins. CCT/TRiC supports proteostasis by promoting correct folding of cytoskeletal proteins such as actin and tubulin, and it influences the maturation of diverse signaling regulators that shape cell growth and stress responses. Through these functions, TCP-1 gamma contributes to pathways governing cytoskeleton organization, cell cycle progression, and protein quality control. Dysregulated chaperonin activity has been associated with altered proteome stability in proliferative and neurodegenerative contexts, making CCT3 a useful node for mechanistic studies of folding-dependent phenotypes.

TCP-1 γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCT3 expression without altering the underlying DNA sequence.

TCP-1 γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCT3 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 CCT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCP-1 γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCT3 locus and enabling the study of TCP-1 γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCP-1 γ pathway restoration in tumor cells with silenced or reduced CCT3 expression.

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