TCP-1 α CRISPR Activation Plasmid (h): sc-402699-ACT

TCP1 encodes TCP-1 alpha, a core subunit of the cytosolic chaperonin-containing TCP1 (CCT/TRiC) complex that folds and stabilizes proteins with complex topologies, including actin and tubulin. By regulating cytoskeletal proteostasis, CCT supports microtubule and actin dynamics, cell-cycle progression, and trafficking processes linked to proteostasis networks. TCP-1 alpha function intersects with stress-response and protein quality-control pathways that shape how cells manage misfolded or aggregation-prone clients. Dysregulated chaperonin activity and altered TCP1 expression have been associated with phenotypes relevant to oncogenic growth, neurodegeneration, and proteostasis imbalance, making it a useful node for mechanistic studies.

TCP-1 α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TCP1 expression without altering the underlying DNA sequence.

TCP-1 α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TCP1 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 TCP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCP-1 α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TCP1 locus and enabling the study of TCP-1 α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCP-1 α pathway restoration in tumor cells with silenced or reduced TCP1 expression.

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