ALKBH5 Double Nickase Plasmid (m): sc-435243-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ALKBH5 Double Nickase Plasmid (m) 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 (m) and ALKBH5 Double Nickase Plasmid (m2) 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 (m) | sc-435243-NIC | 20 µg | $410.00 | |||
ALKBH5 Double Nickase Plasmid (m2) | sc-435243-NIC-2 | 20 µg | $410.00 |
Mouse Alkbh5 encodes ALKBH5, an Fe(II)/2-oxoglutarate–dependent dioxygenase that functions as an RNA N6-methyladenosine (m6A) demethylase, modulating mRNA splicing, export, stability, and translation. By dynamically shaping the epitranscriptomic landscape, ALKBH5 influences gene expression programs linked to cell fate decisions, stress responses, and developmental processes. ALKBH5 activity intersects with m6A writer and reader pathways to regulate transcript-specific outcomes and downstream signaling networks. Dysregulated ALKBH5-mediated RNA methylation has been associated with altered proliferation and differentiation phenotypes in disease-relevant cellular contexts, supporting its use as a node for mechanistic studies of RNA modification biology.
ALKBH5 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Alkbh5 locus in mouse 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.