Rab 9B CRISPR Activation Plasmid (h): sc-412147-ACT

RAB9B encodes the small GTPase Rab 9B, a member of the RAB family that coordinates vesicular trafficking dynamics through regulated GDP/GTP cycling. Rab 9B is implicated in late endosome-to-trans-Golgi network transport and endosomal maturation, processes that influence lysosomal delivery, membrane receptor turnover, and overall endomembrane homeostasis. By shaping intracellular sorting and retrograde trafficking, RAB9B activity can modulate signaling output and organelle function in cell types with high membrane flux. Dysregulation of endolysosomal trafficking pathways is broadly relevant to studies of neurodegeneration, infection biology, and cancer cell adaptation, making RAB9B a useful node for mechanistic interrogation.

Rab 9B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB9B expression without altering the underlying DNA sequence.

Rab 9B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB9B 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 RAB9B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 9B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB9B locus and enabling the study of Rab 9B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 9B pathway restoration in tumor cells with silenced or reduced RAB9B expression.

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