ARHGAP17 CRISPR Activation Plasmid (h): sc-403218-ACT

Human ARHGAP17 encodes a Rho GTPase-activating protein that preferentially downmodulates CDC42 and related Rho-family signaling to coordinate actin cytoskeleton remodeling, cell polarity, and membrane trafficking. By integrating cues that shape junctional organization and motility, ARHGAP17 influences processes such as epithelial barrier regulation, directional migration, and endocytic dynamics. Dysregulated Rho GTPase signaling involving ARHGAP17 has been linked to altered adhesion and invasive phenotypes in cancer biology and to inflammatory signaling contexts where cytoskeletal control impacts immune cell behavior. As a node in Rho GTPase networks, ARHGAP17 is frequently studied to connect upstream receptors to downstream pathways controlling morphology, movement, and tissue organization.

ARHGAP17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARHGAP17 expression without altering the underlying DNA sequence.

ARHGAP17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARHGAP17 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 ARHGAP17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARHGAP17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARHGAP17 locus and enabling the study of ARHGAP17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARHGAP17 pathway restoration in tumor cells with silenced or reduced ARHGAP17 expression.

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