MDA5 Double Nickase Plasmid (h): sc-401962-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MDA5 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
- MDA5 Double Nickase Plasmid (h) and MDA5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting IFIH1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MDA5 Double Nickase Plasmid (h) | sc-401962-NIC | 20 µg | $410.00 | |||
MDA5 Double Nickase Plasmid (h2) | sc-401962-NIC-2 | 20 µg | $410.00 |
IFIH1 encodes melanoma differentiation–associated protein 5 (MDA5), a cytosolic DExD/H-box RNA helicase that detects long double-stranded RNA generated during viral replication. Upon RNA binding, MDA5 signals through MAVS to activate IRF3/IRF7 and NF-κB, driving type I interferon and proinflammatory cytokine programs that shape innate and adaptive immune responses. This pathway intersects with antiviral restriction, apoptosis, and immunometabolic remodeling, and is tightly regulated to prevent aberrant self-RNA sensing. Dysregulated IFIH1/MDA5 activity and interferon signaling have been linked to autoinflammatory and autoimmune phenotypes and can influence inflammatory signaling in infection and cancer-related immune contexts.
MDA5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the IFIH1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within IFIH1. 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 IFIH1 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 IFIH1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.