SCD1 CRISPR Activation Plasmid (m): sc-422809-ACT

Mouse Scd1 encodes stearoyl‑CoA desaturase 1 (SCD1), a microsomal enzyme that introduces a Δ9 double bond into saturated fatty acyl‑CoAs to generate monounsaturated fatty acids such as oleate and palmitoleate. By controlling the saturated/unsaturated lipid balance, SCD1 influences membrane fluidity, triglyceride and cholesterol ester synthesis, lipid droplet formation, and signaling through lipogenic programs including SREBP1‑regulated pathways. Scd1 activity impacts metabolic homeostasis and cellular stress responses, linking altered desaturation capacity to obesity, insulin resistance, hepatic steatosis, inflammation, and context‑dependent effects in cancer cell lipid metabolism. In mouse systems, Scd1 is frequently used to interrogate lipid remodeling, ER stress, and nutrient‑responsive transcriptional networks.

SCD1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Scd1 expression without altering the underlying DNA sequence.

SCD1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Scd1 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 Scd1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Scd1 locus and enabling the study of SCD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCD1 pathway restoration in tumor cells with silenced or reduced Scd1 expression.

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