MITF Double Nickase Plasmid (h): sc-400401-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MITF 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
- MITF Double Nickase Plasmid (h) and MITF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MITF. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MITF Antibody (D-9): sc-515925
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MITF Double Nickase Plasmid (h) | sc-400401-NIC | 20 µg | $410.00 | |||
MITF Double Nickase Plasmid (h2) | sc-400401-NIC-2 | 20 µg | $410.00 |
Microphthalmia-associated transcription factor (MITF) is a basic helix–loop–helix leucine zipper regulator that controls lineage specification and homeostasis in melanocytes and other neural crest–derived cells. It binds E-box motifs to coordinate transcriptional programs governing melanogenesis, pigmentation, lysosome-related organelle biogenesis, proliferation, differentiation, and survival, integrating signals from pathways such as MAPK/ERK, WNT/β-catenin, and cAMP/PKA. MITF also contributes to cellular stress responses and metabolic adaptation through regulation of autophagy and endolysosomal gene networks. Dysregulated MITF activity or expression is linked to pigmentation disorders and is frequently studied in the context of melanoma biology, including tumor cell state switching, invasion, and resistance-associated transcriptional programs.
MITF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MITF locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MITF. 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 MITF 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 MITF-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.