Rab GDI β CRISPR Activation Plasmid (h): sc-404136-ACT

GDI2 encodes Rab GDP dissociation inhibitor beta (Rab GDIβ), a cytosolic chaperone that binds prenylated Rab GTPases and controls their membrane extraction, recycling, and cytosol-to-membrane delivery. By regulating Rab activation cycles, Rab GDIβ supports vesicular trafficking pathways essential for endocytosis, exocytosis, and organelle dynamics, including Golgi and endosomal transport. Altered Rab GTPase homeostasis and trafficking control are linked to dysregulated receptor signaling, antigen presentation, and metabolic adaptation, processes frequently implicated in cancer and neurodegenerative and inflammatory disease biology. As a central coordinator of Rab localization and availability, GDI2 is commonly studied to connect membrane trafficking defects with downstream changes in signaling and cellular phenotype.

Rab GDI β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GDI2 expression without altering the underlying DNA sequence.

Rab GDI β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GDI2 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 GDI2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab GDI β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GDI2 locus and enabling the study of Rab GDI β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab GDI β pathway restoration in tumor cells with silenced or reduced GDI2 expression.

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