TCP-1 ε CRISPR Activation Plasmid (h): sc-402993-ACT

CCT5 encodes the human TCP-1 ε subunit of the TRiC/CCT chaperonin complex, an ATP-dependent folding machine that promotes maturation of nascent and stress-denatured proteins in the cytosol. TRiC supports proteostasis by assisting the folding of key substrates including actin and tubulin, thereby influencing cytoskeletal organization, vesicle trafficking, and cell-cycle progression. Perturbation of chaperonin capacity can alter proteome stability and stress responses, linking CCT5/TCP-1 ε function to pathways that shape cellular fitness under proteotoxic conditions. Dysregulated chaperone activity and impaired folding networks are frequently investigated in the context of neurodegeneration and cancer biology as contributors to protein aggregation, signaling imbalance, and altered proliferation programs.

TCP-1 ε CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCT5 expression without altering the underlying DNA sequence.

TCP-1 ε CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCT5 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 CCT5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCP-1 ε expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCT5 locus and enabling the study of TCP-1 ε-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCP-1 ε pathway restoration in tumor cells with silenced or reduced CCT5 expression.

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