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.
ETO Double Nickase Plasmid (h): sc-401755-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ETO 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
- ETO Double Nickase Plasmid (h) and ETO Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RUNX1T1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ETO Antibody (3H11): sc-134335
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ETO Double Nickase Plasmid (h) | sc-401755-NIC | 20 µg | $410.00 | |||
ETO Double Nickase Plasmid (h2) | sc-401755-NIC-2 | 20 µg | $410.00 |
RUNX1T1 (ETO) encodes a nuclear transcriptional corepressor that partners with DNA-binding transcription factors to modulate gene expression programs controlling hematopoietic differentiation and lineage commitment. ETO contains Nervy homology regions that recruit NCoR/SMRT–HDAC complexes and other chromatin regulators, linking RUNX1T1 to epigenetic silencing, transcriptional repression, and remodeling of enhancer activity. Dysregulated RUNX1T1 function is strongly implicated in leukemogenesis, most notably through RUNX1–RUNX1T1 fusions that perturb RUNX1-dependent transcriptional networks and block myeloid maturation. These pathways make RUNX1T1 a useful target for studying transcriptional repression mechanisms, chromatin-state dynamics, and altered differentiation programs in cancer-relevant cellular models.
ETO Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RUNX1T1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RUNX1T1. 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 RUNX1T1 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 RUNX1T1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.