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.
choactase Double Nickase Plasmid (h): sc-401333-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- choactase 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
- choactase Double Nickase Plasmid (h) and choactase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CHAT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: choactase Antibody (E-7): sc-55557
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
choactase Double Nickase Plasmid (h) | sc-401333-NIC | 20 µg | $410.00 | |||
choactase Double Nickase Plasmid (h2) | sc-401333-NIC-2 | 20 µg | $410.00 |
Human CHAT encodes choline acetyltransferase, the enzyme that catalyzes acetylcholine biosynthesis from choline and acetyl-CoA, supporting cholinergic neurotransmission in central and peripheral nervous systems. CHAT activity links cellular acetyl-CoA metabolism with synaptic vesicle loading and regulated neurotransmitter release, influencing neuronal excitability and neuromuscular signaling. Altered cholinergic tone and CHAT dysregulation have been studied in neurodegeneration, cognitive impairment, and motor neuron and neuromuscular disorders, where acetylcholine availability shapes circuit function and plasticity. As a marker of cholinergic identity, CHAT is widely used to interrogate differentiation programs, synaptic physiology, and neurotransmitter pathway regulation.
choactase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CHAT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CHAT. 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 CHAT 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 CHAT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.