Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
AFP/Alpha fetoprotein Double Nickase Plasmid (h): sc-400355-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AFP/Alpha fetoprotein 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
- AFP/Alpha fetoprotein Double Nickase Plasmid (h) and AFP/Alpha fetoprotein Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AFP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AFP/Alpha fetoprotein Antibody (C3): sc-8399
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AFP/Alpha fetoprotein Double Nickase Plasmid (h) | sc-400355-NIC | 20 µg | $410.00 | |||
AFP/Alpha fetoprotein Double Nickase Plasmid (h2) | sc-400355-NIC-2 | 20 µg | $410.00 |
AFP encodes alpha-fetoprotein, a secreted glycoprotein highly expressed during fetal development and normally silenced postnatally in most tissues. AFP is linked to hepatocyte differentiation programs and fetal-to-adult transcriptional switching, and its re-expression reflects altered gene regulation in liver and germ cell contexts. In cancer biology, AFP is widely used as a molecular marker associated with hepatocellular carcinoma and certain germ cell tumors, supporting studies of tumor cell state, lineage identity, and oncogenic transcriptional networks. As a circulating and tumor-associated antigen, AFP also serves as a readout for investigating secretory pathway dynamics and stress-related changes in protein production.
AFP/Alpha fetoprotein Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AFP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AFP. 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 AFP 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 AFP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.