Rab 1B CRISPR Activation Plasmid (h): sc-402021-ACT

RAB1B encodes the small GTPase Rab 1B, a core regulator of ER-to-Golgi and intra-Golgi vesicular trafficking that coordinates cargo sorting, membrane tethering, and SNARE-dependent fusion. By cycling between GDP- and GTP-bound states, Rab 1B controls early secretory pathway dynamics that influence organelle homeostasis, glycoprotein processing, and secretory proteostasis. Rab1-family signaling interfaces with autophagy and stress-responsive trafficking programs, linking membrane transport to nutrient sensing and quality control. Dysregulated Rab1-mediated trafficking has been associated with cellular phenotypes relevant to neurodegeneration, infection biology, and oncogenic signaling through altered secretion and organelle stress responses.

Rab 1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB1B expression without altering the underlying DNA sequence.

Rab 1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB1B 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 RAB1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB1B locus and enabling the study of Rab 1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 1B pathway restoration in tumor cells with silenced or reduced RAB1B expression.

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