Rab 7L1 CRISPR Activation Plasmid (h): sc-403366-ACT

RAB29 encodes the small GTPase Rab 7L1, a Golgi-associated regulator of membrane trafficking that coordinates vesicle budding, cargo sorting, and endosomal–lysosomal transport dynamics. Rab 7L1 participates in pathways governing retrograde trafficking and Golgi homeostasis, supporting proper localization and turnover of proteins within the secretory and degradative systems. In human cells, RAB29 has been linked to stress-responsive trafficking programs and functional interactions with components of the autophagy–lysosome axis, processes often interrogated in neurodegeneration and inflammatory signaling research. Altered RAB29/Rab 7L1 activity has been studied in the context of Parkinson’s disease–associated networks, including modulation of LRRK2-related trafficking phenotypes, making it relevant for mechanistic studies of organelle quality control.

Rab 7L1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB29 expression without altering the underlying DNA sequence.

Rab 7L1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB29 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 RAB29 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 7L1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB29 locus and enabling the study of Rab 7L1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 7L1 pathway restoration in tumor cells with silenced or reduced RAB29 expression.

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