COPB Double Nickase Plasmid (h): sc-404275-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COPB 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
- COPB Double Nickase Plasmid (h) and COPB Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COPB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: COPB Antibody (D-10): sc-393615
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COPB Double Nickase Plasmid (h) | sc-404275-NIC | 20 µg | $410.00 | |||
COPB Double Nickase Plasmid (h2) | sc-404275-NIC-2 | 20 µg | $410.00 |
COPB1 encodes the COPB subunit of coatomer complex I (COPI), a core component of vesicle coats that mediate retrograde transport from the Golgi to the endoplasmic reticulum and regulate intra-Golgi trafficking. Through its role in cargo selection and vesicle budding, COPB supports maintenance of Golgi architecture, ER homeostasis, and secretory pathway flux, intersecting with proteostasis and stress-response processes such as the unfolded protein response. Perturbation of COPI function can disrupt protein sorting and membrane dynamics, with downstream effects on cell viability and signaling, making COPB1 a useful node for studying organelle communication and membrane trafficking in human cells. Dysregulated intracellular transport pathways are frequently implicated in proliferative and neurodevelopmental disease mechanisms, motivating research into COPB1-dependent trafficking phenotypes.
COPB Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COPB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COPB1. 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 COPB1 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 COPB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.