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Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing
1School of Fisheries, Aquaculture and Aquatic Sciences,Auburn University, 2Department of Animal Wealth Development, Faculty of Veterinary Medicine,Suez Canal University, 3Anatomy and Embryology Department, Faculty of Veterinary Medicine,Cairo University, 4Department of Molecular Physiology and Biophysics,Vanderbilt University, 5Life Science Institute,University of Michigan
Chapters
- 00:00Title
- 01:14Brood Stock Selection and Spawning
- 04:16Sperm Preparation
- 05:33Egg Collection and Fertilization
- 08:07Needle Pulling and Loading
- 10:20Microinjection of Catfish Embryos
- 12:34Results
- 15:24Conclusion
A simple and efficient microinjection protocol for gene editing in channel catfish embryos using the CRISPR/Cas9 system is presented. In this protocol, guide RNAs and Cas9 protein were microinjected into the yolk of one-cell embryos. This protocol has been validated by knocking out two channel catfish immune-related genes.
