ALKBH3 Double Nickase Plasmid (h): sc-406896-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALKBH3 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
- ALKBH3 Double Nickase Plasmid (h) and ALKBH3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALKBH3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ALKBH3 Antibody (B-7): sc-376520
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ALKBH3 Double Nickase Plasmid (h) | sc-406896-NIC | 20 µg | $410.00 | |||
ALKBH3 Double Nickase Plasmid (h2) | sc-406896-NIC-2 | 20 µg | $410.00 |
ALKBH3 (AlkB homolog 3) is a human Fe(II)/2-oxoglutarate–dependent dioxygenase that catalyzes oxidative demethylation of alkylated nucleic acids, contributing to repair of lesions such as 1-methyladenine and 3-methylcytosine in DNA and RNA. This activity supports genome maintenance and RNA quality control, intersecting with DNA damage response pathways and replication-associated stress. Dysregulated ALKBH3 expression and altered alkylation repair capacity have been reported in multiple cancer-related contexts, where dependence on nucleic acid repair and transcriptional fidelity can influence proliferative phenotypes. As a result, ALKBH3 is frequently studied in mechanisms of alkylation damage tolerance, R-loop and transcription-associated instability, and cellular responses to genotoxic stressors.
ALKBH3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALKBH3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALKBH3. 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 ALKBH3 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 ALKBH3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.