FADS1 CRISPR Activation Plasmid (h): sc-404735-ACT

FADS1 encodes fatty acid desaturase 1, an endoplasmic reticulum–localized Δ5 desaturase that catalyzes key steps in the biosynthesis of long-chain polyunsaturated fatty acids, including conversion of dihomo-γ-linolenic acid to arachidonic acid and eicosatetraenoic acid to eicosapentaenoic acid. By shaping cellular pools of ω-6 and ω-3 PUFAs, FADS1 influences membrane composition, lipid mediator production, and downstream signaling linked to inflammation and metabolic homeostasis. Variation or dysregulation of FADS1 has been associated with altered lipid profiles and susceptibility to complex traits involving cardiometabolic and inflammatory biology. In cell and tissue models, FADS1 activity is commonly studied within fatty acid elongation/desaturation networks and pathways governing eicosanoid and specialized pro-resolving mediator precursors.

FADS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FADS1 expression without altering the underlying DNA sequence.

FADS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FADS1 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 FADS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FADS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FADS1 locus and enabling the study of FADS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FADS1 pathway restoration in tumor cells with silenced or reduced FADS1 expression.

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