Rab 5C CRISPR Activation Plasmid (h): sc-403247-ACT

Human RAB5C encodes Rab 5C, a small GTPase that regulates early endosome fusion, receptor-mediated endocytosis, and endosomal trafficking dynamics that shape membrane recycling and signaling duration. By cycling between GDP- and GTP-bound states, Rab 5C coordinates the recruitment of effectors involved in vesicle tethering, cargo sorting, and phosphoinositide remodeling, influencing pathways such as growth factor receptor turnover and nutrient uptake. Rab5 family activity interfaces with cytoskeletal transport and endosome-to-lysosome routing, impacting processes including proliferation, polarity, and responses to extracellular cues. Dysregulated endocytic trafficking and Rab5-dependent signaling have been linked to cancer-associated receptor hyperactivation, neurodegenerative mechanisms tied to vesicle transport deficits, and infectious biology where pathogens exploit host endosomal entry routes.

Rab 5C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB5C expression without altering the underlying DNA sequence.

Rab 5C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB5C 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 RAB5C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 5C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB5C locus and enabling the study of Rab 5C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 5C pathway restoration in tumor cells with silenced or reduced RAB5C expression.

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