Rab 7 CRISPR Activation Plasmid (m): sc-422571-ACT

Rab7 encodes Rab 7, a small GTPase that regulates late endosome maturation, endosome–lysosome trafficking, and lysosomal positioning through coordinated recruitment of effectors that control vesicle motility and membrane fusion. In mouse cells, Rab 7 is central to autophagic flux by promoting autophagosome–lysosome fusion and supporting endocytic cargo degradation, thereby shaping receptor turnover and nutrient-sensing pathways. Disruption of Rab7-dependent trafficking perturbs lysosomal homeostasis and can influence antigen processing, mitochondrial quality control, and neuroimmune signaling. Altered Rab7 activity has been linked to pathological mechanisms involving neurodegeneration, peripheral neuropathy, and infection biology via defects in intracellular trafficking and degradative pathways.

Rab 7 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rab7 expression without altering the underlying DNA sequence.

Rab 7 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Rab7 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 Rab7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Rab7 locus and enabling the study of Rab 7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 7 pathway restoration in tumor cells with silenced or reduced Rab7 expression.

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