RHBDL1 CRISPR Activation Plasmid (h): sc-405676-ACT

RHBDL1 (rhomboid-like 1) encodes an intramembrane serine protease of the rhomboid family that resides in cellular membranes and regulates protein quality control through proteolytic processing of membrane-embedded substrates. By influencing turnover of selected client proteins, RHBDL1 contributes to proteostasis pathways that intersect with ER-associated degradation, membrane protein trafficking, and stress-adaptive signaling. Altered activity of intramembrane proteases has been linked to dysregulated cell signaling and proteome remodeling in cancer and other disorders characterized by chronic cellular stress, making RHBDL1 a relevant target for mechanistic studies. RHBDL1 expression dynamics can also be leveraged to probe how membrane proteolysis shapes pathway responsiveness across diverse human cell contexts.

RHBDL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RHBDL1 expression without altering the underlying DNA sequence.

RHBDL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RHBDL1 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 RHBDL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RHBDL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RHBDL1 locus and enabling the study of RHBDL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RHBDL1 pathway restoration in tumor cells with silenced or reduced RHBDL1 expression.

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