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.
Fatso Double Nickase Plasmid (h): sc-403708-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Fatso 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
- Fatso Double Nickase Plasmid (h) and Fatso Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FTO. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Fatso Antibody (C-3): sc-271713
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Fatso Double Nickase Plasmid (h) | sc-403708-NIC | 20 µg | $410.00 | |||
Fatso Double Nickase Plasmid (h2) | sc-403708-NIC-2 | 20 µg | $410.00 |
Human FTO (Fatso) encodes an Fe(II)/2-oxoglutarate–dependent nucleic acid demethylase that removes N6-methyladenosine (m6A) and related methyl modifications from RNA, thereby shaping mRNA splicing, stability, and translation. Through epitranscriptomic control, FTO influences cellular energy homeostasis, adipogenesis, and nutrient-sensing programs, with downstream effects on pathways linked to mitochondrial function and metabolic stress responses. Genetic variation or dysregulated expression of FTO has been associated with obesity risk and broader metabolic phenotypes, and altered FTO activity is studied in contexts involving growth control and cellular differentiation. These properties make FTO a widely used target for probing RNA methylation–dependent regulation of gene expression in human cells.
Fatso Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FTO locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FTO. 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 FTO 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 FTO-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.