DSCD75 CRISPR Activation Plasmid (h): sc-412188-ACT

Human THEM6 encodes a small membrane-associated protein implicated in endoplasmic reticulum lipid metabolism and phospholipid remodeling, processes that help coordinate membrane biogenesis and cellular stress responses. DSCD75/THEM6 activity has been linked to regulation of fatty acid utilization and homeostatic signaling pathways that couple nutrient availability to organelle function. Perturbation of lipid-handling programs can influence redox balance, proteostasis, and adaptive transcriptional networks, making THEM6 relevant to studies of metabolic reprogramming and cell-state transitions. Altered expression patterns of THEM6 have been reported across disease-associated transcriptomic datasets, supporting its use as a mechanistic node for investigating lipid pathway dysregulation in human cells.

DSCD75 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous THEM6 expression without altering the underlying DNA sequence.

DSCD75 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the THEM6 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 THEM6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DSCD75 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native THEM6 locus and enabling the study of DSCD75-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DSCD75 pathway restoration in tumor cells with silenced or reduced THEM6 expression.

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