CtBP1 CRISPR Activation Plasmid (h): sc-400667-ACT

CTBP1 encodes C-terminal binding protein 1 (CtBP1), a NADH-sensitive transcriptional coregulator that integrates metabolic state with gene expression programs. CtBP1 forms complexes with sequence-specific DNA-binding factors and chromatin modifiers to regulate transcriptional repression and activation, influencing epithelial–mesenchymal transition, cell cycle progression, apoptosis, and differentiation. It participates in chromatin remodeling and transcriptional control pathways by recruiting histone deacetylases and other epigenetic regulators, thereby shaping cellular identity and stress responses. Dysregulated CTBP1 activity has been associated with oncogenic transcriptional networks and altered cell fate decisions in multiple disease-relevant contexts, supporting its use in mechanistic studies of transcription and epigenetics.

CtBP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CTBP1 expression without altering the underlying DNA sequence.

CtBP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CTBP1 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 CTBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CtBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CTBP1 locus and enabling the study of CtBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CtBP1 pathway restoration in tumor cells with silenced or reduced CTBP1 expression.

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