cyclin A Double Nickase Plasmid (h): sc-400119-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- cyclin A 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
- cyclin A Double Nickase Plasmid (h) and cyclin A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CCNA2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: cyclin A Antibody (BF683): sc-239
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
cyclin A Double Nickase Plasmid (h) | sc-400119-NIC | 20 µg | $410.00 | |||
cyclin A Double Nickase Plasmid (h2) | sc-400119-NIC-2 | 20 µg | $410.00 |
CCNA2 encodes cyclin A, a core regulator of cell-cycle progression that coordinates CDK activity during S phase and the G2/M transition. Cyclin A–CDK2 and cyclin A–CDK1 complexes modulate DNA replication origin firing, centrosome duplication, and checkpoint signaling to ensure accurate genome duplication and mitotic entry. CCNA2 expression is tightly controlled by E2F transcriptional programs and ubiquitin-mediated proteolysis via APC/C, linking cyclin A abundance to replication stress and DNA damage responses. Dysregulated CCNA2 activity is frequently associated with altered proliferation control and genome instability, making it a common focus in studies of oncogenic cell-cycle circuitry and chromosomal aberrations.
cyclin A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CCNA2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CCNA2. 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 CCNA2 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 CCNA2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.