EXT1 Double Nickase Plasmid (m): sc-420252-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EXT1 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
- EXT1 Double Nickase Plasmid (m) and EXT1 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Ext1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EXT1 Antibody (A-7): sc-515144
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EXT1 Double Nickase Plasmid (m) | sc-420252-NIC | 20 µg | $410.00 |
Mouse EXT1 encodes exostosin glycosyltransferase 1 (EXT1), an essential component of the EXT1/EXT2 complex that catalyzes heparan sulfate chain elongation in the Golgi apparatus. By controlling heparan sulfate proteoglycan biosynthesis, EXT1 modulates ligand–receptor interactions and gradient formation that influence pathways such as FGF, Wnt, Hedgehog, and BMP signaling. EXT1-dependent glycosaminoglycan structure also affects extracellular matrix organization, cell adhesion, and endocytic trafficking of morphogens and growth factors. Disruption of EXT1 function is associated with aberrant skeletal development and altered tissue patterning, making it a key target for studying heparan sulfate–regulated signaling and matrix biology.
EXT1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Ext1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Ext1. 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 Ext1 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 Ext1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.