Atg12 Double Nickase Plasmid (h): sc-401894-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Atg12 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
- Atg12 Double Nickase Plasmid (h) and Atg12 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATG12. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Atg12 Antibody (C-6): sc-271688
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Atg12 Double Nickase Plasmid (h) | sc-401894-NIC | 20 µg | $410.00 | |||
Atg12 Double Nickase Plasmid (h2) | sc-401894-NIC-2 | 20 µg | $410.00 |
ATG12 encodes Atg12, a ubiquitin-like modifier that is conjugated to ATG5 and associates with ATG16L1 to form the ATG12–ATG5–ATG16L1 complex, a core component of autophagosome biogenesis. This complex supports LC3/ATG8 lipidation and drives expansion of the phagophore during macroautophagy, linking nutrient sensing, organelle quality control, and proteostasis to lysosomal degradation. Through these functions, Atg12 helps shape cellular responses to metabolic stress, oxidative damage, and pathogen challenge, and altered autophagy flux has been implicated in cancer biology, neurodegeneration, and inflammatory disorders. ATG12-dependent pathways are therefore frequently interrogated to study autophagosome formation, selective autophagy, and stress-adaptive signaling networks.
Atg12 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATG12 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATG12. 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 ATG12 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 ATG12-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.