LYPLA2 CRISPR Activation Plasmid (h): sc-405133-ACT

Human LYPLA2 encodes acyl-protein thioesterase 2 (APT2), a serine hydrolase that removes thioester-linked fatty acyl groups from S-palmitoylated proteins, thereby modulating protein membrane association, trafficking, and signaling dynamics. By regulating depalmitoylation cycles, LYPLA2 contributes to lipid-mediated control of subcellular localization and turnover of signaling proteins and can influence endomembrane organization and vesicular transport. This activity connects LYPLA2 to broader lipid homeostasis and post-translational modification networks that shape pathway amplitude and duration in diverse cell types. Dysregulated protein palmitoylation/depalmitoylation has been linked to altered oncogenic signaling, neurobiology, and immune regulation, making LYPLA2 a useful node for mechanistic studies of lipid-driven signaling phenotypes.

LYPLA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LYPLA2 expression without altering the underlying DNA sequence.

LYPLA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LYPLA2 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 LYPLA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LYPLA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LYPLA2 locus and enabling the study of LYPLA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LYPLA2 pathway restoration in tumor cells with silenced or reduced LYPLA2 expression.

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