ACSVL1 CRISPR Activation Plasmid (h): sc-405122-ACT

SLC27A2 encodes acyl‑CoA synthetase very long‑chain 1 (ACSVL1), a fatty acid transport and activation enzyme that converts long‑chain and very‑long‑chain fatty acids into acyl‑CoA thioesters. This reaction supports mitochondrial and peroxisomal lipid utilization, coupling fatty acid uptake to β‑oxidation, lipid remodeling, and energy homeostasis. ACSVL1 activity intersects with pathways governing lipid droplet dynamics, peroxisome function, and metabolic stress responses. Dysregulation of fatty acid activation and very‑long‑chain lipid handling is relevant to studies of metabolic disease mechanisms, inflammatory signaling linked to lipotoxicity, and tissue‑specific lipid metabolism in liver and muscle models.

ACSVL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC27A2 expression without altering the underlying DNA sequence.

ACSVL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC27A2 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 SLC27A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACSVL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC27A2 locus and enabling the study of ACSVL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACSVL1 pathway restoration in tumor cells with silenced or reduced SLC27A2 expression.

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