SKI-1 CRISPR Activation Plasmid (h): sc-402595-ACT

MBTPS1 encodes SKI-1/S1P, a Golgi-resident subtilisin-like proprotein convertase that cleaves membrane-tethered transcription factors to control their activation and nuclear translocation. SKI-1 is a key upstream regulator of sterol and lipid homeostasis through processing of SREBPs, thereby influencing cholesterol and fatty-acid biosynthetic programs and broader metabolic signaling networks. It also participates in ER–Golgi stress-adaptive pathways by modulating the activation of ATF6 and related proteostasis responses. Dysregulation of MBTPS1/SKI-1 activity has been associated with inherited connective tissue and skeletal phenotypes and with cellular states characterized by altered lipid metabolism, making it relevant for mechanistic studies in metabolism, secretory pathway biology, and stress signaling.

SKI-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MBTPS1 expression without altering the underlying DNA sequence.

SKI-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MBTPS1 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 MBTPS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SKI-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MBTPS1 locus and enabling the study of SKI-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SKI-1 pathway restoration in tumor cells with silenced or reduced MBTPS1 expression.

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