HRASLS3 CRISPR Activation Plasmid (h): sc-409231-ACT

PLA2G16 encodes HRASLS3, a membrane-associated phospholipase/acyltransferase that modulates cellular lipid composition by catalyzing phospholipase A1/A2 and lysophospholipid acylation reactions. Through remodeling phosphatidylcholine-derived lipids and generating bioactive fatty acids, HRASLS3 influences membrane dynamics, vesicular trafficking, and lipid-mediated signaling that interfaces with Ras-related pathways and cellular stress responses. Altered PLA2G16/HRASLS3 activity has been linked to dysregulated lipid metabolism and phenotypes relevant to oncogenic transformation, invasion, and inflammatory signaling in multiple model contexts. These properties make HRASLS3 a useful target for mechanistic studies connecting lipid remodeling to proliferation, migration, and metabolic adaptation.

HRASLS3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLA2G16 expression without altering the underlying DNA sequence.

HRASLS3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLA2G16 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 PLA2G16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HRASLS3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLA2G16 locus and enabling the study of HRASLS3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HRASLS3 pathway restoration in tumor cells with silenced or reduced PLA2G16 expression.

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