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.
DNA Ligase IV Double Nickase Plasmid (h): sc-401372-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA Ligase IV 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
- DNA Ligase IV Double Nickase Plasmid (h) and DNA Ligase IV Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting LIG4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DNA Ligase IV Antibody (D-8): sc-271299
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA Ligase IV Double Nickase Plasmid (h) | sc-401372-NIC | 20 µg | $410.00 | |||
DNA Ligase IV Double Nickase Plasmid (h2) | sc-401372-NIC-2 | 20 µg | $410.00 |
LIG4 encodes DNA Ligase IV, an ATP-dependent ligase that seals DNA double-strand break ends during classical non-homologous end joining (c-NHEJ). Acting with XRCC4, XLF/NHEJ1, and the DNA-PK complex, DNA Ligase IV is essential for genome stability, V(D)J recombination, and repair of ionizing radiation–induced lesions. Disruption of LIG4 compromises double-strand break repair fidelity, increasing chromosomal aberrations and sensitivity to genotoxic stress. Pathogenic variants are linked to LIG4 syndrome and broader immunodeficiency and radiosensitivity phenotypes, making this pathway central to studies of DNA damage response and lymphocyte development.
DNA Ligase IV Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the LIG4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within LIG4. 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 LIG4 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 LIG4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.