SOAT1 CRISPR Activation Plasmid (h): sc-417623-ACT

SOAT1 (sterol O-acyltransferase 1; ACAT1) is an endoplasmic reticulum membrane enzyme that catalyzes esterification of free cholesterol to cholesteryl esters, supporting lipid droplet biogenesis and cellular cholesterol homeostasis. By buffering excess sterols, SOAT1 influences membrane composition, lipoprotein metabolism, and sterol-sensitive signaling pathways, including SREBP-regulated lipid programs and inflammatory responses in macrophages. Altered SOAT1 activity has been linked to foam cell formation, neurodegenerative lipid dysregulation, and metabolic rewiring observed in multiple tumor contexts, where cholesteryl ester accumulation can correlate with proliferation and stress adaptation. These biology features make SOAT1 a useful target for studying sterol trafficking, lipid storage, and cholesterol-driven phenotypes in relevant human cell models.

SOAT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SOAT1 expression without altering the underlying DNA sequence.

SOAT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SOAT1 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 SOAT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SOAT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SOAT1 locus and enabling the study of SOAT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SOAT1 pathway restoration in tumor cells with silenced or reduced SOAT1 expression.

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