Atg16 Double Nickase Plasmid (h): sc-404024-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Atg16 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
- Atg16 Double Nickase Plasmid (h) and Atg16 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATG16L1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Atg16 Antibody (E-10): sc-393274
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Atg16 Double Nickase Plasmid (h) | sc-404024-NIC | 20 µg | $410.00 | |||
Atg16 Double Nickase Plasmid (h2) | sc-404024-NIC-2 | 20 µg | $410.00 |
ATG16L1 encodes Atg16, a core component of the ATG12–ATG5–ATG16L1 complex that is required for autophagosome biogenesis and LC3/ATG8 lipidation during macroautophagy. By coordinating phagophore expansion and cargo sequestration, ATG16L1 helps regulate cellular homeostasis, quality control of proteins and organelles, and responses to metabolic and inflammatory stress. ATG16L1-dependent autophagy interfaces with innate immune signaling, including antibacterial xenophagy and control of inflammasome activity, linking this pathway to mucosal barrier and immune regulation. Genetic variation and altered ATG16L1 function have been associated with inflammatory bowel disease susceptibility and dysregulated host–microbe interactions, motivating mechanistic studies in relevant epithelial and immune cell models.
Atg16 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATG16L1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATG16L1. 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 ATG16L1 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 ATG16L1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.