ALKBH5 Double Nickase Plasmid (h): sc-412371-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALKBH5 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
- ALKBH5 Double Nickase Plasmid (h) and ALKBH5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALKBH5. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ALKBH5 Double Nickase Plasmid (h) | sc-412371-NIC | 20 µg | $410.00 | |||
ALKBH5 Double Nickase Plasmid (h2) | sc-412371-NIC-2 | 20 µg | $410.00 |
Human ALKBH5 encodes an Fe(II)/2-oxoglutarate–dependent dioxygenase that functions as an N6-methyladenosine (m6A) RNA demethylase, shaping epitranscriptomic regulation of mRNA metabolism. By modulating m6A levels, ALKBH5 influences RNA splicing, nuclear export, translation, and stability, thereby impacting programs that govern cell-cycle progression, stress adaptation, and differentiation. ALKBH5 activity intersects with broader RNA-processing pathways and can alter the expression dynamics of oncogenic and developmental transcripts. Dysregulated ALKBH5-dependent m6A turnover has been implicated in disease-relevant phenotypes including aberrant proliferation, altered stem-like states, and disrupted immune and metabolic signaling networks.
ALKBH5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALKBH5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALKBH5. 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 ALKBH5 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 ALKBH5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.