Rab 15 CRISPR Activation Plasmid (h): sc-415722-ACT

RAB15 encodes Rab 15, a small Ras-related GTPase that regulates endosomal membrane trafficking by cycling between GDP- and GTP-bound states and coordinating vesicle budding, transport, and fusion. Rab 15 activity has been linked to receptor endocytosis and sorting decisions that shape signaling amplitude and nutrient uptake, intersecting with Rab-dependent endocytic recycling and lysosomal targeting processes. Through its role in endosomal dynamics, Rab 15 can influence pathways governing cellular metabolism, membrane receptor homeostasis, and compartment-specific signal transduction. Altered expression or function of endocytic Rab proteins, including Rab 15, is frequently investigated in contexts such as oncogenic signaling rewiring, neurobiology, and immune cell regulation where trafficking-dependent phenotypes are prominent.

Rab 15 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB15 expression without altering the underlying DNA sequence.

Rab 15 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB15 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 RAB15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 15 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB15 locus and enabling the study of Rab 15-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 15 pathway restoration in tumor cells with silenced or reduced RAB15 expression.

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