DOT1L1 Double Nickase Plasmid (h): sc-403286-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DOT1L1 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
- DOT1L1 Double Nickase Plasmid (h) and DOT1L1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DOT1L. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DOT1L1 Antibody (E-5): sc-374317
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DOT1L1 Double Nickase Plasmid (h) | sc-403286-NIC | 20 µg | $410.00 | |||
DOT1L1 Double Nickase Plasmid (h2) | sc-403286-NIC-2 | 20 µg | $410.00 |
DOT1L encodes DOT1L1, a lysine methyltransferase that catalyzes mono-, di-, and tri-methylation of histone H3 Lys79, a chromatin mark linked to active transcription. Through regulation of gene expression programs, DOT1L influences cell-cycle progression, DNA damage responses, hematopoietic differentiation, and developmental transcriptional networks. DOT1L-dependent H3K79 methylation interfaces with epigenetic control of RNA polymerase II elongation and chromatin accessibility, shaping lineage-specific transcriptional states. Dysregulated DOT1L activity and altered H3K79 methylation patterns are implicated in oncogenic transcriptional programs, including those associated with MLL-rearranged leukemias, supporting its relevance in epigenetics and cancer biology research.
DOT1L1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DOT1L locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DOT1L. 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 DOT1L 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 DOT1L-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.