MBD4 Double Nickase Plasmid (h): sc-403156-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MBD4 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
- MBD4 Double Nickase Plasmid (h) and MBD4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MBD4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MBD4 Antibody (D-6): sc-271530
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MBD4 Double Nickase Plasmid (h) | sc-403156-NIC | 20 µg | $410.00 |
MBD4 (methyl-CpG binding domain protein 4) is a DNA glycosylase that recognizes G:T and G:U mismatches arising from deamination of methylated cytosines and initiates base excision repair to preserve CpG integrity. Through its methyl-CpG binding domain and interaction with the mismatch repair machinery, MBD4 links epigenetic context to genome surveillance and helps limit mutation accumulation at methylated loci. Disruption of MBD4 function is associated with elevated C>T transition signatures and microsatellite instability phenotypes, connecting this pathway to mechanisms of genome instability observed in multiple cancer types. As a result, MBD4 is widely studied in DNA repair, mutation processes, and epigenome-maintenance research.
MBD4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MBD4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MBD4. 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 MBD4 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 MBD4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.