RAPGEF6 CRISPR Activation Plasmid (h): sc-403816-ACT

RAPGEF6 (also known as PDZ-GEF2) encodes a Rap-specific guanine nucleotide exchange factor that activates RAP1 and RAP2 GTPases to coordinate integrin inside-out signaling, cytoskeletal remodeling, and cell–cell junction dynamics. Through regulation of Rap-dependent pathways, RAPGEF6 contributes to control of cell adhesion, migration, and polarity across multiple tissue contexts. Its activity interfaces with signaling networks downstream of receptors that influence MAPK and small GTPase circuitry, shaping trafficking and membrane-proximal signaling. Dysregulated Rap signaling and adhesion programs linked to RAPGEF6 are relevant to studies of invasive cell behavior, immune cell trafficking, and other disease-associated alterations in tissue organization.

RAPGEF6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAPGEF6 expression without altering the underlying DNA sequence.

RAPGEF6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAPGEF6 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 RAPGEF6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RAPGEF6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAPGEF6 locus and enabling the study of RAPGEF6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RAPGEF6 pathway restoration in tumor cells with silenced or reduced RAPGEF6 expression.

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