Rab 10 CRISPR Activation Plasmid (m): sc-422551-ACT

Rab10 encodes Rab 10, a small Ras-related GTPase that coordinates membrane trafficking by regulating vesicle budding, transport, tethering, and fusion between endosomes, recycling pathways, and the trans-Golgi network. In mouse cells, Rab 10 activity supports polarized exocytosis, GLUT4 vesicle trafficking, autophagosome maturation, and receptor recycling, linking it to PI3K–AKT signaling, endosomal sorting, and cytoskeletal dynamics. Perturbation of Rab10-dependent trafficking has been associated with altered insulin responsiveness, immune cell function, and neuronal homeostasis, making Rab10 a useful node for studying metabolic and neurodegeneration-relevant pathways in vivo and in cultured models.

Rab 10 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rab10 expression without altering the underlying DNA sequence.

Rab 10 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Rab10 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 Rab10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Rab10 locus and enabling the study of Rab 10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 10 pathway restoration in tumor cells with silenced or reduced Rab10 expression.

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