Rab 2A CRISPR Activation Plasmid (h): sc-403154-ACT

Human RAB2A encodes the small GTPase Rab 2A, a core regulator of ER-to-Golgi and intra-Golgi vesicular trafficking that coordinates cargo sorting, vesicle tethering, and fusion events through cycling between GDP- and GTP-bound states. Rab 2A supports secretory pathway homeostasis and contributes to organelle organization, including Golgi integrity and membrane dynamics that influence protein processing and delivery. Through its role in membrane trafficking, RAB2A intersects with pathways controlling proteostasis, receptor turnover, and stress responses linked to cellular growth and survival. Dysregulated Rab GTPase signaling and trafficking programs, including RAB2A-associated networks, are frequently studied in the context of oncogenic signaling, neurobiology, and pathogen–host interactions where vesicle transport capacity can shape disease-relevant phenotypes.

Rab 2A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB2A expression without altering the underlying DNA sequence.

Rab 2A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB2A 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 RAB2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 2A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB2A locus and enabling the study of Rab 2A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 2A pathway restoration in tumor cells with silenced or reduced RAB2A expression.

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