PTPLAD2 CRISPR Activation Plasmid (h2): sc-417162-ACT-2

Human HACD4 (encoding PTPLAD2) is a putative 3-hydroxyacyl-CoA dehydratase–like enzyme implicated in the elongation of very-long-chain fatty acids, linking it to lipid metabolic homeostasis, membrane biogenesis, and endoplasmic reticulum–associated processes. Through its predicted role in fatty acid remodeling, HACD4/PTPLAD2 is relevant to pathways governing phospholipid composition, organelle integrity, and cellular stress responses that influence proliferation and differentiation. Altered regulation of lipid-elongation networks and PTPLAD2-associated signatures have been reported in contexts of metabolic dysfunction and cancer-related biology, supporting its utility as a mechanistic node for studying lipid-driven phenotypes. Gene editing of HACD4 enables functional dissection of VLCFA metabolism, interrogation of downstream signaling and transcriptomic adaptations, and development of cell models for lipidomic, ER stress, and membrane dynamics research.

PTPLAD2 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous HACD4 expression without altering the underlying DNA sequence.

PTPLAD2 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HACD4 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 HACD4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PTPLAD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HACD4 locus and enabling the study of PTPLAD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PTPLAD2 pathway restoration in tumor cells with silenced or reduced HACD4 expression.

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