Rab 11B CRISPR Activation Plasmid (h): sc-402256-ACT

RAB11B encodes the small GTPase Rab 11B, a key regulator of recycling endosome dynamics that coordinates membrane trafficking from endosomes back to the plasma membrane and through the trans-Golgi network. By cycling between GDP- and GTP-bound states, Rab 11B helps control vesicle budding, transport, and fusion events that shape receptor recycling, integrin turnover, and polarized cargo delivery. Rab11-family signaling intersects with cytoskeletal remodeling and cell migration programs through effector interactions that influence actin organization and membrane composition. Dysregulated endocytic recycling and vesicle transport associated with RAB11B activity has been linked to altered signaling homeostasis and phenotypes relevant to cancer cell invasion, neurobiology, and broader trafficking-related disease mechanisms.

Rab 11B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB11B expression without altering the underlying DNA sequence.

Rab 11B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB11B 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 RAB11B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 11B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB11B locus and enabling the study of Rab 11B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 11B pathway restoration in tumor cells with silenced or reduced RAB11B expression.

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