CD39L4 CRISPR Activation Plasmid (h): sc-410720-ACT

Human ENTPD5 encodes CD39L4, an endoplasmic reticulum–localized ectonucleoside triphosphate diphosphohydrolase that hydrolyzes UDP and GDP to monophosphates, supporting nucleotide sugar homeostasis and glycoprotein quality control. By sustaining the UDP-glucose–dependent calnexin/calreticulin folding cycle, CD39L4 influences ER protein processing, N-glycosylation, and proteostasis programs linked to secretion and membrane receptor maturation. ENTPD5 activity intersects with ER stress signaling and metabolic rewiring through effects on protein folding demand and glycan-dependent trafficking. Dysregulated ENTPD5 expression has been associated with altered glycosylation phenotypes and proliferation-related pathways in cancer biology, making it relevant for studies of ER homeostasis and oncogenic signaling networks.

CD39L4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ENTPD5 expression without altering the underlying DNA sequence.

CD39L4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ENTPD5 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 ENTPD5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD39L4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ENTPD5 locus and enabling the study of CD39L4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD39L4 pathway restoration in tumor cells with silenced or reduced ENTPD5 expression.

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