connexin 50 Double Nickase Plasmid (h): sc-404332-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- connexin 50 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 50 Double Nickase Plasmid (h) and connexin 50 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GJA8. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: connexin 50 Antibody (B-11): sc-373801
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
connexin 50 Double Nickase Plasmid (h) | sc-404332-NIC | 20 µg | $410.00 | |||
connexin 50 Double Nickase Plasmid (h2) | sc-404332-NIC-2 | 20 µg | $410.00 |
GJA8 encodes connexin 50, a gap junction protein that assembles into hexameric connexons to mediate intercellular exchange of ions and small metabolites. In the ocular lens, connexin 50 supports epithelial–fiber cell communication, contributing to lens homeostasis, transparency, and coordinated differentiation during development. Its activity intersects with processes regulating membrane permeability, calcium signaling, and maintenance of the lens microcirculation system that sustains avascular tissue physiology. Disruption of GJA8-dependent coupling is associated with altered lens fiber organization and cataract-related phenotypes, making it a useful entry point for studying gap junction biology in ocular models.
connexin 50 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GJA8 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GJA8. 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 GJA8 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 GJA8-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.