TTC36 CRISPR Activation Plasmid (m): sc-431423-ACT

Mouse Ttc36 encodes TTC36, a tetratricopeptide repeat (TPR)-containing protein predicted to function as a scaffold for multiprotein complexes that coordinate protein–protein interactions. TPR-domain proteins commonly influence proteostasis, intracellular trafficking, and signaling by modulating chaperone engagement and the assembly of regulatory complexes. Although TTC36 remains incompletely characterized, expression- and genetics-driven studies can clarify its contribution to cell-state control programs such as stress response, differentiation, and tissue homeostasis. Dysregulation of TPR-mediated complex formation is frequently implicated in disease-associated phenotypes, making Ttc36 a useful target for mechanistic exploration in mouse models.

TTC36 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ttc36 expression without altering the underlying DNA sequence.

TTC36 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ttc36 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 Ttc36 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TTC36 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ttc36 locus and enabling the study of TTC36-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TTC36 pathway restoration in tumor cells with silenced or reduced Ttc36 expression.

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