Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Dnmt2 Double Nickase Plasmid (h): sc-402709-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Dnmt2 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
- Dnmt2 Double Nickase Plasmid (h) and Dnmt2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TRDMT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Dnmt2 Antibody (D-9): sc-365001
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Dnmt2 Double Nickase Plasmid (h) | sc-402709-NIC | 20 µg | $410.00 | |||
Dnmt2 Double Nickase Plasmid (h2) | sc-402709-NIC-2 | 20 µg | $410.00 |
TRDMT1 encodes Dnmt2, an evolutionarily conserved RNA cytosine-5 methyltransferase that primarily modifies specific tRNAs (e.g., m5C at tRNA Asp-GTC) rather than genomic DNA. This activity contributes to tRNA stability, translational fidelity, and cellular adaptation to stress, linking TRDMT1 to RNA epigenetic regulation and proteostasis. Dnmt2-mediated tRNA methylation has been associated with control of small RNA biogenesis and stress-response pathways that influence cell survival and differentiation. Altered TRDMT1 function or expression has been reported in multiple cancer contexts and in models of neurodevelopmental and metabolic dysregulation, motivating mechanistic studies of RNA methylation in disease-relevant phenotypes.
Dnmt2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TRDMT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TRDMT1. 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 TRDMT1 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 TRDMT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.