Rab 38 CRISPR Activation Plasmid (h): sc-404742-ACT

RAB38 encodes Rab 38, a small Rab family GTPase that regulates vesicular trafficking and organelle biogenesis, with prominent roles in endosome-to-lysosome transport and the maturation of lysosome-related organelles. Rab 38 coordinates cargo sorting and membrane dynamics through interactions with tethering and effector complexes, influencing protein delivery to melanosomes and other specialized secretory compartments. Perturbation of RAB38-dependent trafficking can alter pigment granule formation, lysosomal function, and secretion phenotypes, linking this pathway to cellular homeostasis in pigment cells and immune-related secretory programs. As a node in intracellular transport networks, Rab 38 is relevant for mechanistic studies of membrane trafficking defects observed across diverse disease-associated cellular states.

Rab 38 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB38 expression without altering the underlying DNA sequence.

Rab 38 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB38 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 RAB38 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 38 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB38 locus and enabling the study of Rab 38-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 38 pathway restoration in tumor cells with silenced or reduced RAB38 expression.

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