Cdx2 CRISPR Activation Plasmid (h): sc-401220-ACT

CDX2 encodes the caudal type homeobox transcription factor Cdx2, a key regulator of intestinal development, epithelial differentiation, and maintenance of anterior–posterior patterning programs. Cdx2 controls gene networks involved in cell fate specification, barrier function, and lineage-restricted transcriptional programs in gastrointestinal tissues, integrating with Wnt/β-catenin, BMP, and Notch-associated signaling contexts. Dysregulated CDX2 expression or altered CDX2-dependent transcriptional circuitry has been associated with aberrant differentiation states and gastrointestinal pathology, and it is widely used as a marker of intestinal lineage identity in human model systems. As a sequence-specific DNA-binding factor, Cdx2 provides a tractable node for dissecting enhancer usage, chromatin state transitions, and transcriptional network remodeling during epithelial plasticity.

Cdx2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDX2 expression without altering the underlying DNA sequence.

Cdx2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDX2 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 CDX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdx2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDX2 locus and enabling the study of Cdx2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdx2 pathway restoration in tumor cells with silenced or reduced CDX2 expression.

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