MLH1 Double Nickase Plasmid (m): sc-421660-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MLH1 Double Nickase Plasmid (m) 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
- MLH1 Double Nickase Plasmid (m) and MLH1 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Mlh1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MLH1 Antibody (B-12): sc-271978
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MLH1 Double Nickase Plasmid (m) | sc-421660-NIC | 20 µg | $410.00 | |||
MLH1 Double Nickase Plasmid (m2) | sc-421660-NIC-2 | 20 µg | $410.00 |
Mlh1 encodes MLH1, a core component of the DNA mismatch repair (MMR) machinery that safeguards genome integrity by correcting base–base mismatches and insertion–deletion loops arising during DNA replication. MLH1 functions within MutL complexes, coordinating with MutS homologs to couple mismatch recognition to excision and resynthesis, and it also participates in DNA damage signaling and the processing of recombination intermediates. In mouse cells, impaired MLH1 activity increases replication-associated mutagenesis and microsatellite instability, linking MMR defects to genomic instability phenotypes widely used in cancer biology and DNA repair research. Accordingly, Mlh1 is frequently studied in pathways governing mutation accumulation, checkpoint responses, and genome maintenance.
MLH1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Mlh1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Mlh1. 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 Mlh1 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 Mlh1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.