Rab 32 CRISPR Activation Plasmid (h): sc-403733-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rab 32 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Rab 32 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Rab 32 CRISPR Activation Plasmid (h) and Rab 32 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RAB32 transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rab 32 Antibody (D-11): sc-377472
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rab 32 CRISPR Activation Plasmid (h) | sc-403733-ACT | 20 µg | $397.00 |
RAB32 encodes Rab 32, a small GTPase of the RAB family that regulates intracellular membrane trafficking, including transport between endosomes, lysosome-related organelles, and mitochondria-associated compartments. Rab 32 participates in vesicle budding, cargo sorting, and organelle homeostasis, linking Rab-dependent trafficking to mitochondrial dynamics and cellular stress responses. Through its role in trafficking and organelle quality control, altered RAB32 activity has been associated with dysregulated inflammation and aberrant cellular metabolism in disease-relevant contexts. These functions make RAB32 a useful node for studying vesicular transport pathways, organelle biogenesis, and signaling networks that depend on compartmentalized protein localization.
Rab 32 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB32 expression without altering the underlying DNA sequence.
Rab 32 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB32 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 RAB32 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 32 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB32 locus and enabling the study of Rab 32-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 32 pathway restoration in tumor cells with silenced or reduced RAB32 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.