connexin 30.3 Double Nickase Plasmid (h): sc-409363-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- connexin 30.3 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 30.3 Double Nickase Plasmid (h) and connexin 30.3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GJB4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: connexin 30.3 Antibody (F6): sc-81802
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
connexin 30.3 Double Nickase Plasmid (h) | sc-409363-NIC | 20 µg | $410.00 |
GJB4 encodes connexin 30.3, a gap junction channel subunit that assembles into connexons to mediate direct intercellular exchange of ions and small metabolites. Through these junctions, connexin 30.3 contributes to tissue-level coordination of membrane potential, calcium signaling, and metabolic coupling, supporting epithelial differentiation and homeostasis. In the epidermis, connexin-mediated communication intersects with keratinocyte proliferation and barrier maintenance pathways. Dysregulated GJB4 function and altered gap junction connectivity have been associated with inherited skin disorders characterized by aberrant cornification and impaired epithelial integrity.
connexin 30.3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GJB4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GJB4. 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 GJB4 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 GJB4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.