Histone H2A.X Double Nickase Plasmid (h): sc-400538-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Histone H2A.X 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
- Histone H2A.X Double Nickase Plasmid (h) and Histone H2A.X Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting H2AFX. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Histone H2A.X Antibody (938CT5.1.1): sc-517336
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Histone H2A.X Double Nickase Plasmid (h) | sc-400538-NIC | 20 µg | $410.00 | |||
Histone H2A.X Double Nickase Plasmid (h2) | sc-400538-NIC-2 | 20 µg | $410.00 |
H2AFX encodes histone H2A.X, a variant of histone H2A that becomes rapidly phosphorylated at Ser139 (γH2AX) at sites of DNA double-strand breaks. This modification functions as an early chromatin-based signal that recruits and organizes DNA damage response factors, coordinating checkpoint activation and repair pathway choice across homologous recombination and non-homologous end joining. H2A.X-dependent signaling interfaces with ATM/ATR-mediated stress responses, replication fork stability, and chromatin remodeling that collectively preserve genome integrity. Dysregulation of H2A.X signaling or compromised γH2AX dynamics is widely used as a molecular readout of genotoxic stress and is relevant to studies of genomic instability, cancer biology, and radiosensitivity.
Histone H2A.X Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the H2AFX locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within H2AFX. 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 H2AFX 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 H2AFX-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.