GPR116 CRISPR Activation Plasmid (h): sc-408424-ACT

ADGRF5 (GPR116) is an adhesion G protein–coupled receptor with a large extracellular N-terminus and GPCR proteolysis site that links extracellular matrix and cell–cell interactions to intracellular signaling. It is expressed in epithelial and endothelial compartments, including the lung, where it contributes to tissue homeostasis and barrier-related processes through GPCR-mediated pathways such as cAMP/PKA and Rho GTPase signaling. GPR116 has been implicated in regulation of surfactant balance and alveolar function, and altered expression has been associated with inflammatory remodeling and fibrotic responses in respiratory tissues. Dysregulation of ADGRF5 has also been reported across multiple cancer transcriptomic datasets, supporting its use in studies of tumor–microenvironment interactions, adhesion signaling, and cell migration.

GPR116 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADGRF5 expression without altering the underlying DNA sequence.

GPR116 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADGRF5 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 ADGRF5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR116 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADGRF5 locus and enabling the study of GPR116-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR116 pathway restoration in tumor cells with silenced or reduced ADGRF5 expression.

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