GPRC5D CRISPR Activation Plasmid (h): sc-403114-ACT

GPRC5D (G protein-coupled receptor class C group 5 member D) encodes an orphan GPCR-like membrane protein enriched in epithelial contexts and linked to differentiation-associated signaling. As a retinoic acid–responsive family member, GPRC5D is studied for its connections to transcriptional programs that influence cell state, membrane receptor signaling, and microenvironmental interactions. Altered GPRC5D expression has been reported in hematologic malignancy research, particularly plasma cell biology, and is also examined in solid tumor and immune-oncology datasets. These features make GPRC5D a useful target for investigating receptor-linked regulatory networks and lineage-associated phenotypes in human cells.

GPRC5D CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPRC5D expression without altering the underlying DNA sequence.

GPRC5D CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPRC5D 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 GPRC5D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPRC5D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPRC5D locus and enabling the study of GPRC5D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPRC5D pathway restoration in tumor cells with silenced or reduced GPRC5D expression.

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