METTL3 Double Nickase Plasmid (h): sc-404029-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- METTL3 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
- METTL3 Double Nickase Plasmid (h) and METTL3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting METTL3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: METTL3 Antibody (D-9): sc-518180
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
METTL3 Double Nickase Plasmid (h) | sc-404029-NIC | 20 µg | $410.00 | |||
METTL3 Double Nickase Plasmid (h2) | sc-404029-NIC-2 | 20 µg | $410.00 |
METTL3 encodes the catalytic core of the mRNA N6-methyladenosine (m6A) methyltransferase complex, partnering with METTL14 and cofactors such as WTAP to install m6A marks on coding and noncoding RNAs. This RNA modification influences transcript stability, splicing, nuclear export, and translation, thereby shaping programs linked to cell-cycle control, differentiation, stress responses, and innate immune signaling. METTL3-dependent m6A regulation intersects with RNA metabolism pathways and epitranscriptomic control of gene expression, including modulation of signaling outputs such as MAPK, PI3K–AKT, and DNA damage response networks through altered mRNA fate. Dysregulated METTL3 activity has been associated with diverse disease-relevant phenotypes in cancer biology, hematopoiesis, and inflammatory contexts, making it a frequent target in mechanistic studies of RNA-based regulation.
METTL3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the METTL3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within METTL3. 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 METTL3 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 METTL3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.