ChoK CRISPR Activation Plasmid (h): sc-403793-ACT

Human CHKA encodes choline kinase alpha (ChoK), the ATP-dependent enzyme that catalyzes phosphorylation of choline to phosphocholine, a rate-limiting step in the Kennedy pathway for phosphatidylcholine biosynthesis. Through control of phosphocholine pools and membrane phospholipid production, ChoK supports cellular membrane biogenesis, lipid signaling, and metabolic remodeling associated with proliferation and stress responses. CHKA activity intersects with oncogenic signaling programs and altered choline metabolism observed across multiple tumor types, and it has also been implicated in inflammatory and neurodegenerative contexts through broader phospholipid homeostasis. As a metabolic node linking nutrient utilization to membrane dynamics, CHKA is frequently studied in pathways governing cell growth, migration, and organelle function.

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

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

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