APG5/ATG5 Double Nickase Plasmid (h): sc-416847-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- APG5/ATG5 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
- APG5/ATG5 Double Nickase Plasmid (h) and APG5/ATG5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATG5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: APG5/ATG5 Antibody (C-1): sc-133158
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
APG5/ATG5 Double Nickase Plasmid (h) | sc-416847-NIC | 20 µg | $410.00 | |||
APG5/ATG5 Double Nickase Plasmid (h2) | sc-416847-NIC-2 | 20 µg | $410.00 |
ATG5 (APG5) is an essential autophagy factor that conjugates with ATG12 and associates with ATG16L1 to form a core complex required for autophagosome membrane elongation and LC3 lipidation. Through these functions, ATG5 supports basal and stress-induced macroautophagy, mitophagy, and the clearance of protein aggregates and damaged organelles, integrating nutrient-sensing pathways such as mTOR and AMPK with lysosomal turnover. Altered ATG5 activity has been linked to dysregulated proteostasis, inflammation, and susceptibility to neurodegenerative, metabolic, infectious, and cancer-associated phenotypes, making it a frequent target in studies of cell survival and immune signaling. In human cells, ATG5 perturbation is commonly used to dissect autophagy-dependent versus autophagy-independent mechanisms in stress responses and intracellular quality control.
APG5/ATG5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATG5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATG5. 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 ATG5 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 ATG5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.