Suez Canal University

2 ARTICLES PUBLISHED IN JoVE

JoVE Core

Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing

Ahmed Elaswad^*1,2, Karim Khalil^*1,3, David Cline¹, Patrick Page-McCaw⁴, Wenbiao Chen⁴, Maximilian Michel⁵, Roger Cone⁵, Rex Dunham¹

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

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.

Biology

Single-Cell Suspension Preparation from Nile Tilapia Intestine for Single-Cell Sequencing

Peibo Wang^*1, Yongcan Zhou^*1,3, Bei Wang², Ahmed Elaswad⁴, Shifeng Wang^1,3, Weiliang Guo^1,3, Dongdong Zhang^1,3

¹State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, ²Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, College of Fishery, Guangdong Ocean University, ³Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, ⁴Department of Animal Wealth Development, Faculty of Veterinary Medicine, Suez Canal University

Here, we demonstrate the preparation of a high-quality single-cell suspension of tilapia intestine for single-cell sequencing.