TM6SF2 CRISPR Activation Plasmid (h): sc-408066-ACT

Human TM6SF2 (transmembrane 6 superfamily member 2) encodes an endoplasmic reticulum–associated multi-pass membrane protein implicated in hepatic lipid handling and very-low-density lipoprotein (VLDL) secretion. TM6SF2 modulates triglyceride-rich lipoprotein assembly and influences intracellular triglyceride accumulation, linking it to metabolic pathways governing lipid droplet dynamics and ER homeostasis. Genetic variation and altered expression of TM6SF2 have been associated with dyslipidemia and susceptibility to fatty liver phenotypes, supporting its relevance in studies of metabolic liver disease. As a regulatory node in hepatocyte lipid flux, TM6SF2 is frequently examined in contexts spanning lipid metabolism, inflammation-associated stress responses, and cardiometabolic risk mechanisms.

TM6SF2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TM6SF2 expression without altering the underlying DNA sequence.

TM6SF2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TM6SF2 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 TM6SF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TM6SF2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TM6SF2 locus and enabling the study of TM6SF2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TM6SF2 pathway restoration in tumor cells with silenced or reduced TM6SF2 expression.

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