Ret Double Nickase Plasmid (m): sc-422654-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ret 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
- Ret Double Nickase Plasmid (m) and Ret Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Ret. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ret Antibody (6E4C4): sc-101423
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ret Double Nickase Plasmid (m) | sc-422654-NIC | 20 µg | $410.00 |
Ret encodes a receptor tyrosine kinase that functions as a signaling hub for GDNF family ligands via GFRα co-receptors, coordinating neuronal and renal developmental programs in mouse. Upon activation, RET engages canonical MAPK/ERK, PI3K–AKT, and PLCγ pathways to regulate cell survival, differentiation, migration, and axon guidance. Ret activity is central to neural crest-derived lineages and enteric nervous system formation, and its dysregulation is linked to neurodevelopmental defects and oncogenic signaling contexts. As a pathway node, Ret is widely studied to dissect growth factor signaling, lineage specification, and receptor-driven transcriptional responses.
Ret Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Ret locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Ret. 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 Ret 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 Ret-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.