ROM-K Double Nickase Plasmid (h): sc-404241-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ROM-K Double Nickase Plasmid (h) 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
- ROM-K Double Nickase Plasmid (h) and ROM-K Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KCNJ1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ROM-K Antibody (D-3): sc-393189
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ROM-K Double Nickase Plasmid (h) | sc-404241-NIC | 20 µg | $410.00 | |||
ROM-K Double Nickase Plasmid (h2) | sc-404241-NIC-2 | 20 µg | $410.00 |
KCNJ1 encodes ROM-K (Kir1.1), an inwardly rectifying potassium channel that mediates K⁺ recycling across the apical membrane of renal tubular epithelia, supporting sodium reabsorption and potassium homeostasis. ROM-K activity couples to transepithelial ion transport processes in the thick ascending limb and collecting duct, influencing membrane potential and driving electrogenic transport. The channel participates in renal electrolyte handling pathways that intersect with aldosterone-regulated transport programs and cellular volume/osmotic responses. Dysregulation or loss of KCNJ1 function is linked to inherited salt-wasting tubulopathies characterized by impaired renal K⁺ conservation and altered acid–base balance, making it a useful target for mechanistic studies of ion transport disorders.
ROM-K Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KCNJ1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KCNJ1. 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 KCNJ1 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 KCNJ1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.