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 I Double Nickase Plasmid (h): sc-402830-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA Ligase I 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 I Double Nickase Plasmid (h) and DNA Ligase I Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting LIG1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DNA Ligase I Antibody (C-5): sc-271678
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA Ligase I Double Nickase Plasmid (h) | sc-402830-NIC | 20 µg | $410.00 | |||
DNA Ligase I Double Nickase Plasmid (h2) | sc-402830-NIC-2 | 20 µg | $410.00 |
Human LIG1 encodes DNA ligase I, an ATP-dependent ligase that seals single-strand breaks by joining Okazaki fragments during lagging-strand synthesis and completing repair-associated DNA nick ligation. DNA ligase I functions at the interface of replication and multiple DNA repair pathways, including long-patch base excision repair and resolution of DNA strand discontinuities that arise during DNA damage processing. Proper LIG1 activity supports genome stability, replication fork progression, and cell-cycle fidelity, and its disruption is associated with elevated replication stress and mutagenesis. Altered ligation capacity has been implicated in genomic instability phenotypes relevant to cancer biology and inherited DNA repair deficiency syndromes, making LIG1 a useful node for studying DNA maintenance mechanisms.
DNA Ligase I Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the LIG1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within LIG1. 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 LIG1 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 LIG1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.