SREBP-2 CRISPR Activation Plasmid (h): sc-400575-ACT

SREBF2 encodes sterol regulatory element-binding protein 2 (SREBP-2), a membrane-tethered transcription factor that is proteolytically activated under low sterol conditions to induce genes controlling cholesterol biosynthesis and uptake. In concert with SCAP and INSIG proteins, SREBP-2 regulates the mevalonate pathway and LDL receptor–mediated cholesterol import, coordinating lipid homeostasis with ER stress and metabolic cues. Altered SREBF2/SREBP-2 activity has been implicated in dyslipidemia and metabolic syndrome–related phenotypes, and has also been linked to lipid rewiring that supports proliferation and inflammatory signaling in disease contexts. As a central node in sterol sensing and transcriptional control, SREBF2 is widely studied for its role in metabolic regulation, membrane composition, and crosstalk with autophagy and innate immune pathways.

SREBP-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SREBF2 expression without altering the underlying DNA sequence.

SREBP-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SREBF2 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 SREBF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SREBP-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SREBF2 locus and enabling the study of SREBP-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SREBP-2 pathway restoration in tumor cells with silenced or reduced SREBF2 expression.

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