Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Rab 27a Double Nickase Plasmid (m): sc-419219-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rab 27a Double Nickase Plasmid (m) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Rab 27a Double Nickase Plasmid (m) and Rab 27a Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Rab27a. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rab 27a Antibody (E-8): sc-74586
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rab 27a Double Nickase Plasmid (m) | sc-419219-NIC | 20 µg | $410.00 | |||
Rab 27a Double Nickase Plasmid (m2) | sc-419219-NIC-2 | 20 µg | $410.00 |
Rab27a encodes the small GTPase Rab 27a, a membrane trafficking regulator that controls docking and fusion of secretory vesicles with the plasma membrane. In mouse cells, Rab 27a coordinates regulated exocytosis through interactions with effectors such as melanophilin, Slp/Exophilin proteins, and Munc13-4, supporting processes that include melanosome transport, cytotoxic granule release, and platelet dense granule secretion. This pathway intersects with actin-dependent vesicle transport, lysosome-related organelle biogenesis, and immune synapse function. Dysregulated Rab 27a activity has been linked to pigmentation phenotypes and immune dysfunction in genetic models, making it a useful node for studying vesicle dynamics and organelle homeostasis.
Rab 27a Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Rab27a locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Rab27a. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt Rab27a function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of Rab27a-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.