SCD4 CRISPR Activation Plasmid (m): sc-435929-ACT

Mouse Scd4 encodes stearoyl-CoA desaturase 4 (SCD4), an endoplasmic reticulum-associated enzyme that introduces a cis double bond into saturated fatty acyl-CoAs to generate monounsaturated fatty acids. By shaping the cellular balance of saturated versus unsaturated lipids, SCD4 influences membrane fluidity, lipid droplet biogenesis, and the availability of substrates for triglyceride, phospholipid, and cholesterol ester synthesis. Scd4 activity intersects with lipogenic transcriptional programs regulated by SREBP/INSIG signaling and integrates with metabolic pathways controlling fatty acid oxidation and energy homeostasis. Altered desaturase-dependent lipid remodeling is relevant to studies of metabolic dysfunction and inflammation, where shifts in lipid saturation can modulate ER stress responses and downstream signaling outputs.

SCD4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Scd4 expression without altering the underlying DNA sequence.

SCD4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Scd4 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 Scd4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCD4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Scd4 locus and enabling the study of SCD4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCD4 pathway restoration in tumor cells with silenced or reduced Scd4 expression.

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