HMGCR CRISPR Activation Plasmid (m): sc-420877-ACT

Mouse Hmgcr encodes 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), the rate-limiting enzyme of the mevalonate pathway that converts HMG-CoA to mevalonate and controls cellular cholesterol and isoprenoid biosynthesis. By regulating sterol availability, HMGCR influences membrane composition, lipoprotein metabolism, and feedback control of lipid homeostasis through SREBP-dependent transcriptional programs and ER-associated degradation. Flux through the mevalonate pathway also supports protein prenylation, impacting small GTPase signaling, vesicular trafficking, and cell growth regulation. Altered HMGCR activity and mevalonate-pathway output are widely used as experimental entry points for studying dyslipidemia-associated mechanisms, metabolic inflammation, and lipid-dependent modulation of cell proliferation and differentiation.

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

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

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