Pancreatic Lipase CRISPR Activation Plasmid (h): sc-402264-ACT

PNLIP encodes pancreatic lipase, a secreted digestive hydrolase that catalyzes the hydrolysis of dietary triacylglycerols into monoacylglycerols and free fatty acids in the intestinal lumen. Acting at the lipid–water interface in concert with colipase and bile salts, PNLIP supports intestinal lipid absorption and downstream metabolic handling of fatty acids and fat-soluble nutrients. Altered PNLIP expression or activity has been associated with exocrine pancreatic dysfunction and disorders of fat digestion, and it is frequently used as a functional marker in studies of pancreatic acinar biology. Because lipid digestion influences systemic energy balance and nutrient signaling, PNLIP is also relevant to research on metabolic phenotypes linked to pancreatic function.

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

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

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