ACSVL4 CRISPR Activation Plasmid (h): sc-402011-ACT

SLC27A4 encodes ACSVL4 (FATP4), a very long-chain acyl-CoA synthetase that couples long-chain and very long-chain fatty acid uptake to intracellular activation, thereby channeling lipids into β-oxidation, complex lipid biosynthesis, and membrane remodeling. By generating fatty acyl-CoAs, ACSVL4 influences epithelial lipid homeostasis and barrier-associated processes, and intersects with metabolic programs that regulate cellular stress responses and differentiation. Dysregulated SLC27A4 activity has been linked to defects in epidermal lipid processing and altered fatty acid handling, making it relevant for studies of skin biology, lipid metabolism, and metabolic reprogramming in disease-associated contexts.

ACSVL4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC27A4 expression without altering the underlying DNA sequence.

ACSVL4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC27A4 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 SLC27A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACSVL4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC27A4 locus and enabling the study of ACSVL4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACSVL4 pathway restoration in tumor cells with silenced or reduced SLC27A4 expression.

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