PRA1 CRISPR Activation Plasmid (h): sc-404295-ACT

Human RABAC1 encodes PRA1, a prenylated Rab acceptor that functions as a regulator of Rab GTPase trafficking and membrane dynamics. PRA1 participates in endomembrane transport by modulating Rab localization and cycling, influencing ER–Golgi trafficking, vesicle tethering, and cargo sorting across secretory and endocytic routes. Through these roles, RABAC1 connects to proteostasis and cellular stress responses that depend on coordinated vesicular transport. Dysregulated Rab-mediated trafficking and secretory pathway control are frequently implicated in phenotypes relevant to cancer cell biology and neurodegeneration research, making PRA1 a useful node for mechanistic studies.

PRA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RABAC1 expression without altering the underlying DNA sequence.

PRA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RABAC1 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 RABAC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RABAC1 locus and enabling the study of PRA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRA1 pathway restoration in tumor cells with silenced or reduced RABAC1 expression.

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