ALKB Double Nickase Plasmid (h): sc-408412-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALKB 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
- ALKB Double Nickase Plasmid (h) and ALKB Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALKBH1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ALKB Antibody (F-4): sc-374301
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ALKB Double Nickase Plasmid (h) | sc-408412-NIC | 20 µg | $410.00 | |||
ALKB Double Nickase Plasmid (h2) | sc-408412-NIC-2 | 20 µg | $410.00 |
ALKBH1 encodes a Fe(II)/2-oxoglutarate–dependent dioxygenase implicated in oxidative demethylation of nucleic acids and regulation of chromatin-associated processes. The protein has been linked to DNA repair and replication-associated stress responses, and it contributes to maintenance of genome stability through modulation of base modifications. ALKBH1 activity intersects with pathways governing epigenetic regulation, mitochondrial and nuclear nucleic acid metabolism, and cellular adaptation to genotoxic stress. Dysregulation of ALKBH1 expression or function has been associated with altered proliferation programs and tumor-relevant phenotypes, making it a useful target for mechanistic studies in cancer biology and DNA damage signaling.
ALKB Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALKBH1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALKBH1. 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 ALKBH1 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 ALKBH1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.