Fatty Acid Synthase CRISPR Activation Plasmid (h): sc-400440-ACT

Human FASN encodes fatty acid synthase, a multifunctional cytosolic enzyme complex that catalyzes de novo synthesis of long-chain saturated fatty acids, primarily palmitate, from acetyl-CoA and malonyl-CoA using NADPH. This activity integrates with acetyl-CoA metabolism, citrate shuttling, and NADPH-generating pathways to support membrane biogenesis, protein palmitoylation, and lipid-derived signaling. FASN regulation is closely linked to nutrient sensing and lipogenic transcriptional programs, including SREBP-dependent control, and it contributes to cellular responses to hypoxia and oxidative stress through lipid remodeling. Altered FASN expression and lipogenesis are frequently studied in contexts of metabolic dysfunction and proliferative disease biology, where lipid availability can influence growth, stress tolerance, and inflammatory signaling.

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

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

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