connexin 46 Double Nickase Plasmid (h): sc-403947-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- connexin 46 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
- connexin 46 Double Nickase Plasmid (h) and connexin 46 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GJA3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: connexin 46 Antibody (C-3): sc-365394
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
connexin 46 Double Nickase Plasmid (h) | sc-403947-NIC | 20 µg | $410.00 | |||
connexin 46 Double Nickase Plasmid (h2) | sc-403947-NIC-2 | 20 µg | $410.00 |
GJA3 encodes connexin 46, a lens-enriched gap junction protein that assembles into hemichannels and intercellular channels to mediate direct cell–cell transfer of ions and small metabolites. Connexin 46 supports lens fiber cell differentiation and homeostasis by contributing to the lens microcirculation system, coordinating redox balance, and maintaining ionic and osmotic equilibrium. Through regulation of junctional communication and hemichannel permeability, it influences calcium handling, ATP signaling, and cellular stress responses in avascular ocular tissue. Disruption of GJA3 function is associated with inherited cataract phenotypes, making it a relevant target for studying lens transparency mechanisms and protein quality control in the ocular environment.
connexin 46 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GJA3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GJA3. 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 GJA3 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 GJA3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.