Rab 33B CRISPR Activation Plasmid (h): sc-404070-ACT

RAB33B encodes Rab 33B, a small GTPase of the Rab family that regulates membrane trafficking events linked to Golgi and autophagy-associated compartments. By cycling between GDP- and GTP-bound states, Rab 33B coordinates vesicle budding, transport, and tethering/fusion to support cargo sorting and organelle homeostasis. RAB33B-dependent trafficking intersects with macroautophagy pathways and intracellular protein turnover, processes frequently studied in the context of neurodegeneration, infection biology, and cancer cell stress responses. Disruption or dysregulation of Rab-mediated transport networks can perturb secretory pathway flux and autophagic dynamics, providing mechanistic entry points for disease-relevant cell biology research.

Rab 33B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB33B expression without altering the underlying DNA sequence.

Rab 33B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB33B 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 RAB33B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 33B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB33B locus and enabling the study of Rab 33B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 33B pathway restoration in tumor cells with silenced or reduced RAB33B expression.

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