Rab 20 CRISPR Activation Plasmid (h): sc-412563-ACT

RAB20 encodes Rab20, a small GTPase that regulates vesicular trafficking within the endocytic system, coordinating membrane transport events that influence receptor recycling, endosome maturation, and compartmentalized signaling. By cycling between GTP- and GDP-bound states, Rab20 helps organize vesicle budding, motility, and fusion through interactions with effectors that couple trafficking to cytoskeletal dynamics and organelle function. Altered Rab GTPase network activity, including RAB20 dysregulation, has been associated with perturbations in cellular homeostasis such as metabolic adaptation, inflammatory signaling, and oncogenic phenotypes in multiple experimental contexts. As a result, RAB20 is frequently studied in pathways linking intracellular transport to stress responses and disease-relevant cell states.

Rab 20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB20 expression without altering the underlying DNA sequence.

Rab 20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB20 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 RAB20 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB20 locus and enabling the study of Rab 20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 20 pathway restoration in tumor cells with silenced or reduced RAB20 expression.

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