ALR Double Nickase Plasmid (h): sc-403367-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALR 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
- ALR Double Nickase Plasmid (h) and ALR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GFER. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ALR Antibody (E-7): sc-365885
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ALR Double Nickase Plasmid (h) | sc-403367-NIC | 20 µg | $410.00 | |||
ALR Double Nickase Plasmid (h2) | sc-403367-NIC-2 | 20 µg | $410.00 |
Human GFER encodes augmenter of liver regeneration (ALR), a sulfhydryl oxidase that localizes primarily to the mitochondrial intermembrane space and supports oxidative protein folding via the MIA40–GFER disulfide relay. By promoting import and maturation of cysteine-rich mitochondrial proteins, ALR contributes to respiratory chain maintenance, redox homeostasis, and mitochondrial integrity, influencing energy metabolism and stress responses. GFER activity intersects with apoptotic regulation and cellular proliferation through mitochondrial signaling and reactive oxygen species handling. Dysregulation of ALR/GFER has been linked to mitochondrial dysfunction phenotypes and is frequently investigated in contexts of hepatic biology, neurodevelopment, and disorders associated with impaired proteostasis and redox imbalance.
ALR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GFER locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GFER. 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 GFER 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 GFER-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.