53BP1 Double Nickase Plasmid (h): sc-400321-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 53BP1 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
- 53BP1 Double Nickase Plasmid (h) and 53BP1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TP53BP1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: 53BP1 Antibody (E-10): sc-515841
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
53BP1 Double Nickase Plasmid (h) | sc-400321-NIC | 20 µg | $410.00 | |||
53BP1 Double Nickase Plasmid (h2) | sc-400321-NIC-2 | 20 µg | $410.00 |
TP53BP1 encodes 53BP1, a chromatin-associated DNA damage response factor that accumulates at double-strand breaks through recognition of histone marks and cooperation with upstream sensors such as ATM. 53BP1 promotes non-homologous end joining and constrains DNA end resection, thereby influencing repair pathway choice in opposition to BRCA1-dependent homologous recombination. Through these activities, it supports genome stability, S-phase progression, and checkpoint signaling, and its dysregulation is implicated in mutational burden and chromosomal rearrangements observed in multiple cancer contexts. TP53BP1 is also widely used as a functional readout for DNA damage foci formation and repair competence in studies of replication stress and genotoxic responses.
53BP1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TP53BP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TP53BP1. 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 TP53BP1 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 TP53BP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.