CHPT1 CRISPR Activation Plasmid (h2): sc-412745-ACT-2

Human CHPT1 (choline phosphotransferase 1) encodes an endoplasmic reticulum–associated enzyme that catalyzes the terminal step of phosphatidylcholine biosynthesis in the Kennedy pathway by transferring phosphocholine from CDP-choline to diacylglycerol. By regulating cellular phosphatidylcholine production, CHPT1 contributes to membrane biogenesis, lipid droplet dynamics, and maintenance of organelle membrane composition, with downstream effects on secretory trafficking and stress responses linked to lipid homeostasis. Perturbation of phosphatidylcholine metabolism is implicated in metabolic dysfunction and oncogenic phenotypes through altered membrane signaling platforms and lipid remodeling. CHPT1 is therefore a useful target for gene editing studies probing phospholipid flux, ER membrane biology, and genotype-to-phenotype relationships in lipid-associated disease models.

CHPT1 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous CHPT1 expression without altering the underlying DNA sequence.

CHPT1 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHPT1 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 CHPT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHPT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHPT1 locus and enabling the study of CHPT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHPT1 pathway restoration in tumor cells with silenced or reduced CHPT1 expression.

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