CDD CRISPR Activation Plasmid (h): sc-403915-ACT

Human CDA encodes cytidine deaminase (CDD), a zinc-dependent enzyme that catalyzes deamination of cytidine and deoxycytidine to uridine and deoxyuridine, respectively, supporting pyrimidine salvage and nucleoside homeostasis. By regulating intracellular nucleoside pools, CDD influences DNA replication and repair capacity and can modulate cellular responses to nucleoside analog exposure through metabolic inactivation. CDA activity intersects with broader nucleotide metabolism networks that shape proliferation, replication stress tolerance, and mitochondrial–nuclear balance of dNTP availability. Dysregulated CDA/CDD expression or activity has been associated with altered nucleoside metabolism phenotypes relevant to cancer biology, immune cell function, and inherited variation in drug metabolism pathways, making it a useful node for mechanistic studies.

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

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

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