cytohesin-2 CRISPR Activation Plasmid (h): sc-403025-ACT

Human CYTH2 encodes cytohesin-2, an ARF guanine nucleotide exchange factor that activates ARF small GTPases to coordinate membrane trafficking, phosphoinositide-dependent signaling, and actin cytoskeleton remodeling. Through its Sec7 catalytic domain and pleckstrin homology–mediated membrane recruitment, cytohesin-2 supports receptor recycling, integrin-dependent adhesion, and downstream pathways including PI3K/AKT and MAPK that shape cell motility and survival responses. CYTH2 activity has been studied in contexts of immune cell activation and inflammatory signaling, and dysregulated cytohesin/ARF axis function is frequently examined for its contribution to aberrant migration and growth control in disease-associated cellular phenotypes. These properties make CYTH2 a useful target for dissecting endocytic regulation and signaling network plasticity in mechanistic cell biology.

cytohesin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYTH2 expression without altering the underlying DNA sequence.

cytohesin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYTH2 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 CYTH2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous cytohesin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYTH2 locus and enabling the study of cytohesin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of cytohesin-2 pathway restoration in tumor cells with silenced or reduced CYTH2 expression.

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