SCAP CRISPR Activation Plasmid (h): sc-401474-ACT

Human SCAP (SREBF chaperone) is an endoplasmic reticulum membrane protein that functions as a sterol sensor and escort factor for SREBP transcription factors, enabling regulated trafficking to the Golgi for proteolytic activation. Through this SCAP–SREBP axis, SCAP controls transcriptional programs governing cholesterol and fatty acid biosynthesis, membrane lipid homeostasis, and adaptive responses to nutrient and sterol availability. SCAP activity interfaces with INSIG-mediated retention, ER-to-Golgi transport, and feedback regulation of lipogenic gene expression. Dysregulation of SCAP-dependent signaling has been linked to altered lipid metabolism observed in metabolic disease and to lipid reprogramming that supports growth and stress adaptation in multiple cancer contexts.

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

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

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