Autotaxin CRISPR Activation Plasmid (h): sc-401889-ACT

ENPP2 encodes autotaxin, a secreted lysophospholipase D that converts lysophosphatidylcholine into lysophosphatidic acid (LPA), a potent lipid mediator controlling cell migration, survival, cytoskeletal remodeling, and vascular permeability. Autotaxin-driven LPA signaling engages GPCR-dependent pathways including Rho/ROCK, PI3K–AKT, and MAPK/ERK, linking extracellular lipid metabolism to transcriptional programs and motility. ENPP2 activity is regulated within inflammatory and stromal microenvironments and contributes to processes such as wound repair, fibrosis, and angiogenesis. Dysregulated autotaxin–LPA axis biology is implicated in cancer invasion and metastasis, chronic inflammation, and fibrotic disease mechanisms, making ENPP2 a useful target for pathway dissection in human cell models.

Autotaxin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ENPP2 expression without altering the underlying DNA sequence.

Autotaxin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ENPP2 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 ENPP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Autotaxin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ENPP2 locus and enabling the study of Autotaxin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Autotaxin pathway restoration in tumor cells with silenced or reduced ENPP2 expression.

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