CtBP2 CRISPR Activation Plasmid (h2): sc-401865-ACT-2

Human CTBP2 encodes the C-terminal binding protein 2 (CtBP2), a NADH-sensitive transcriptional corepressor that modulates gene expression by partnering with DNA-binding factors and recruiting chromatin-modifying complexes. CtBP2 regulates programs linked to cell fate determination, epithelial–mesenchymal transition, cell-cycle control, and apoptosis, integrating metabolic state with transcriptional outputs across developmental and stress-responsive pathways. Dysregulated CTBP2 activity has been associated with altered oncogenic transcriptional networks and aberrant differentiation in multiple cancer contexts, making it relevant for mechanistic studies of tumor progression and metastasis. Gene editing of CTBP2 supports functional interrogation of corepressor-dependent chromatin regulation, mapping of transcriptional circuits, and validation of pathway dependencies using reporter assays, omics readouts, and perturbation screens.

CtBP2 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous CTBP2 expression without altering the underlying DNA sequence.

CtBP2 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CTBP2 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 CTBP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CtBP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CTBP2 locus and enabling the study of CtBP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CtBP2 pathway restoration in tumor cells with silenced or reduced CTBP2 expression.

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