| Chapter title |
Gene Editing in Channel Catfish via Double Electroporation of Zinc-Finger Nucleases
|
|---|---|
| Chapter number | 15 |
| Book title |
Zinc Finger Proteins
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-8799-3_15 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8798-6, 978-1-4939-8799-3
|
| Authors |
Rex A. Dunham, Ahmed Elaswad, Zhenkui Qin, Dunham, Rex A., Elaswad, Ahmed, Qin, Zhenkui |
| Abstract |
The traditional approach for gene editing with zinc-finger nucleases (ZFNs) in fish has been microinjection of mRNA. Here, we develop and describe an alternative protocol in which ZFN plasmids are electroporated to channel catfish, Ictalurus punctatus, sperm, and embryos. Briefly, plasmids were propagated to supply a sufficient quantity for electroporation. Sperm cells were prepared in saline solution, electroporated with plasmids, and then used for fertilization. Embryos were incubated with the plasmids before performing electroporation just prior to first cell division. Plasmids were then transcribed and translated by embryonic cells to produce ZFNs for gene editing, resulting in mutated fry. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 10 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
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| Student > Master | 1 | 10% |
| Student > Bachelor | 1 | 10% |
| Researcher | 1 | 10% |
| Other | 1 | 10% |
| Other | 0 | 0% |
| Unknown | 4 | 40% |
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|---|---|---|
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| Biochemistry, Genetics and Molecular Biology | 1 | 10% |
| Agricultural and Biological Sciences | 1 | 10% |
| Immunology and Microbiology | 1 | 10% |
| Psychology | 1 | 10% |
| Other | 1 | 10% |
| Unknown | 4 | 40% |