Ku-86 Double Nickase Plasmid (h): sc-400549-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ku-86 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
- Ku-86 Double Nickase Plasmid (h) and Ku-86 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting XRCC5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ku86 Antibody (B-1): sc-5280
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ku-86 Double Nickase Plasmid (h) | sc-400549-NIC | 20 µg | $410.00 | |||
Ku-86 Double Nickase Plasmid (h2) | sc-400549-NIC-2 | 20 µg | $410.00 |
XRCC5 encodes Ku-86 (Ku80), a core subunit of the Ku heterodimer that recognizes DNA double-strand breaks and initiates canonical non-homologous end joining (c-NHEJ). By binding broken DNA ends and recruiting DNA-PKcs to form the DNA-PK complex, Ku-86 coordinates end processing and ligation to preserve genome integrity during replication stress and after genotoxic insults. Ku-86 function intersects with V(D)J recombination, class switch recombination, telomere maintenance, and DNA damage signaling checkpoints. Dysregulation of XRCC5-dependent repair has been associated with genomic instability phenotypes and altered sensitivity to DNA-damaging agents, supporting its frequent study in cancer biology, immunology, and aging-related genome maintenance.
Ku-86 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the XRCC5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within XRCC5. 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 XRCC5 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 XRCC5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.