Histone H3.3A Double Nickase Plasmid (h): sc-416802-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Histone H3.3A 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 H3.3A Double Nickase Plasmid (h) and Histone H3.3A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting H3F3A. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Histone H3.3A Double Nickase Plasmid (h) | sc-416802-NIC | 20 µg | $410.00 | |||
Histone H3.3A Double Nickase Plasmid (h2) | sc-416802-NIC-2 | 20 µg | $410.00 |
H3F3A encodes the replication-independent histone variant H3.3A, a core component of nucleosomes that is deposited at actively transcribed genes, regulatory elements, telomeres, and sites of DNA damage. Through variant-specific incorporation, H3.3 helps shape chromatin accessibility and coordinates transcriptional regulation, enhancer activity, and epigenetic memory, while also influencing DNA repair and genome stability. H3.3A participates in chromatin remodeling and histone modification networks that interface with processes such as replication stress responses and cell fate programs. Recurrent H3F3A alterations and dysregulated H3.3 deposition have been linked to aberrant chromatin states in cancer and developmental contexts, making this locus a focal point for mechanistic studies of epigenetic dysregulation.
Histone H3.3A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the H3F3A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within H3F3A. 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 H3F3A 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 H3F3A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.