α/β-dystroglycan Double Nickase Plasmid (h): sc-417547-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α/β-dystroglycan 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
- α/β-dystroglycan Double Nickase Plasmid (h) and α/β-dystroglycan Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DAG1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: α-dystroglycan Antibody (IIH6): sc-53987
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
α/β-dystroglycan Double Nickase Plasmid (h) | sc-417547-NIC | 20 µg | $410.00 | |||
α/β-dystroglycan Double Nickase Plasmid (h2) | sc-417547-NIC-2 | 20 µg | $410.00 |
DAG1 encodes dystroglycan, a laminin-binding cell surface receptor that is post-translationally processed into α-dystroglycan and β-dystroglycan and assembled into the dystrophin–glycoprotein complex. This complex couples the extracellular matrix to the actin cytoskeleton, supporting basement membrane organization, membrane stability, and mechanotransduction in skeletal muscle and other tissues. Dystroglycan function is tightly linked to glycosylation-dependent ligand binding and signaling at adhesion sites, integrating extracellular matrix cues with cytoskeletal remodeling. Disruption of DAG1 expression or dystroglycan processing/glycosylation is associated with dystroglycanopathies and related neuromuscular phenotypes, making it a key node for studying cell–matrix interactions and tissue integrity.
α/β-dystroglycan Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DAG1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DAG1. 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 DAG1 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 DAG1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.