AFG3L2 Double Nickase Plasmid (h): sc-404565-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AFG3L2 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
- AFG3L2 Double Nickase Plasmid (h) and AFG3L2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AFG3L2. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AFG3L2 Double Nickase Plasmid (h) | sc-404565-NIC | 20 µg | $410.00 |
AFG3L2 encodes a mitochondrial inner membrane AAA+ metalloprotease that forms the m-AAA protease complex, supporting ATP-dependent protein quality control within the intermembrane-facing side of the inner membrane. By regulating turnover and maturation of respiratory chain components and mitochondrial ribosome-related substrates, AFG3L2 contributes to oxidative phosphorylation efficiency, mitochondrial proteostasis, and stress-adaptive signaling. Loss or dysfunction of AFG3L2 perturbs mitochondrial dynamics and bioenergetics, promoting accumulation of misfolded proteins and impaired organelle homeostasis. Variants in AFG3L2 are associated with neurodegenerative phenotypes, linking mitochondrial protease activity to neuronal vulnerability and axonal maintenance.
AFG3L2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AFG3L2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AFG3L2. 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 AFG3L2 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 AFG3L2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.