Atg4b Double Nickase Plasmid (h): sc-404173-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Atg4b 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
- Atg4b Double Nickase Plasmid (h) and Atg4b Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATG4B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Atg4b Antibody (231CT21.1.7): sc-517310
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Atg4b Double Nickase Plasmid (h) | sc-404173-NIC | 20 µg | $410.00 | |||
Atg4b Double Nickase Plasmid (h2) | sc-404173-NIC-2 | 20 µg | $410.00 |
ATG4B encodes Atg4b, a cysteine protease that processes ATG8 family proteins, including LC3 and GABARAP, by priming them for lipidation and recycling them through delipidation during autophagosome maturation. Through these steps, Atg4b regulates autophagic flux, mitophagy, and the turnover of damaged proteins and organelles, linking it to cellular stress adaptation and metabolic homeostasis. Autophagy pathway modulation by ATG4B intersects with mTOR and AMPK signaling and influences antigen presentation and innate immune responses. Dysregulated ATG4B activity and altered autophagy dynamics have been associated with cancer biology, neurodegeneration, and inflammatory phenotypes, making ATG4B a useful node for mechanistic studies of proteostasis.
Atg4b Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATG4B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATG4B. 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 ATG4B 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 ATG4B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.