Rab 14 CRISPR Activation Plasmid (h): sc-402109-ACT

RAB14 encodes Rab 14, a small GTPase of the RAS superfamily that coordinates membrane trafficking by cycling between GDP- and GTP-bound states to control vesicle budding, transport, and fusion. Rab 14 regulates endocytic recycling and biosynthetic transport between the trans-Golgi network, early/recycling endosomes, and the plasma membrane, influencing cargo sorting, receptor turnover, and polarized trafficking. Through these processes, RAB14 contributes to maintenance of cell polarity, nutrient transporter distribution, and signal transduction dynamics. Altered Rab14-regulated trafficking has been linked in the literature to dysregulated proliferation and migration programs observed across multiple disease contexts, making it a useful node for mechanistic studies of vesicle transport–dependent phenotypes.

Rab 14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB14 expression without altering the underlying DNA sequence.

Rab 14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB14 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 RAB14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB14 locus and enabling the study of Rab 14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 14 pathway restoration in tumor cells with silenced or reduced RAB14 expression.

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