Ly-GDI/RhoGDI2/ARHGDIB CRISPR Activation Plasmid (h): sc-402326-ACT

ARHGDIB encodes Ly-GDI (RhoGDI2), a hematopoietic-enriched Rho GDP dissociation inhibitor that binds prenylated Rho family GTPases and controls their membrane cycling and activation state. By modulating RAC1, RHOA, and CDC42 signaling, ARHGDIB regulates actin cytoskeleton dynamics, cell adhesion, chemotaxis, and phagocyte functional responses central to innate immune signaling. This pathway interfaces with integrin-mediated motility and inflammatory signaling networks that coordinate leukocyte trafficking and tissue surveillance. Dysregulated ARHGDIB expression has been linked to altered immune cell migration and inflammatory phenotypes and is frequently studied in the context of hematologic malignancy biology and tumor microenvironment interactions.

Ly-GDI/RhoGDI2/ARHGDIB CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARHGDIB expression without altering the underlying DNA sequence.

Ly-GDI/RhoGDI2/ARHGDIB CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARHGDIB 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 ARHGDIB transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ly-GDI/RhoGDI2/ARHGDIB expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARHGDIB locus and enabling the study of Ly-GDI/RhoGDI2/ARHGDIB-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ly-GDI/RhoGDI2/ARHGDIB pathway restoration in tumor cells with silenced or reduced ARHGDIB expression.

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