Nicotinic Acetylcholine Receptor epsilon/CHRNE Double Nickase Plasmid (h): sc-404780-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nicotinic Acetylcholine Receptor epsilon/CHRNE 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
- Nicotinic Acetylcholine Receptor epsilon/CHRNE Double Nickase Plasmid (h) and Nicotinic Acetylcholine Receptor epsilon/CHRNE Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CHRNE. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Nicotinic Acetylcholine Receptor epsilon/CHRNE Antibody (B-11): sc-376747
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nicotinic Acetylcholine Receptor epsilon/CHRNE Double Nickase Plasmid (h) | sc-404780-NIC | 20 µg | $410.00 | |||
Nicotinic Acetylcholine Receptor epsilon/CHRNE Double Nickase Plasmid (h2) | sc-404780-NIC-2 | 20 µg | $410.00 |
CHRNE encodes the epsilon subunit of the muscle-type nicotinic acetylcholine receptor (nAChR), a ligand-gated cation channel that is essential for fast synaptic transmission at the neuromuscular junction. Incorporation of the epsilon subunit into the adult nAChR pentamer supports acetylcholine-dependent channel opening, sodium influx, and endplate depolarization that initiates excitation–contraction coupling. CHRNE function integrates with agrin–LRP4–MuSK signaling, acetylcholinesterase-regulated neurotransmitter clearance, and activity-dependent receptor clustering and turnover at the postsynaptic membrane. Pathogenic CHRNE variants are linked to congenital myasthenic syndromes characterized by impaired neuromuscular transmission and altered receptor expression or gating kinetics.
Nicotinic Acetylcholine Receptor epsilon/CHRNE Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CHRNE locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CHRNE. 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 CHRNE 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 CHRNE-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.