dCK CRISPR Activation Plasmid (h): sc-417715-ACT

Human DCK encodes deoxycytidine kinase (dCK), a rate-limiting enzyme in the nucleoside salvage pathway that phosphorylates deoxycytidine and related deoxynucleosides to their monophosphate forms, supporting balanced dNTP pools for DNA replication and repair. dCK activity links nucleotide metabolism to cell-cycle progression, replication stress responses, and genome maintenance, particularly in proliferative tissues. Dysregulated DCK expression or activity has been associated with altered nucleotide homeostasis and metabolic vulnerabilities observed across hematologic and solid tumor contexts, and it is frequently studied in relation to cellular differentiation and stress adaptation. As a central node in nucleoside metabolism, DCK is a useful target for probing pathway cross-talk with mitochondrial function, redox state, and DNA damage signaling.

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

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

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