Cairo University

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

Mohamed Salem^1,2, Michal Bernach¹, Krzysztof Bajdzienko¹, Patrick Giavalisco¹

¹Max Planck Institute of Molecular Plant Physiology, Golm, Germany, ²Department of Pharmacognosy, Faculty of Pharmacy, Cairo University

A protocol for comprehensive extraction of lipids, metabolites and proteins from biological tissues using one sample is presented.

Genetics

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.

Genetics

Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice

Hirofumi Nishizono^*1,2,3, Mohamed Darwish^*4,5, Hideki Uosaki^6,7, Nanami Masuyama^8,9,10, Motoaki Seki^8,11, Hiroyuki Abe³, Nozomu Yachie^8,9,10,12,13, Ryohei Yasuda¹

¹Max Planck Florida Institute for Neuroscience, ²Life Science Research Center, University of Toyama, ³Department of Biochemical Engineering, Graduate School of Science and Engineering, Yamagata University, ⁴Graduate School of Innovative Life Science, University of Toyama, ⁵Department of Biochemistry, Faculty of Pharmacy, Cairo University, ⁶Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, ⁷Division of Stem Cell Research and Drug Development, Center for Development of Advanced Medical Technology, Jichi Medical University, ⁸Synthetic Biology Division, Research Center for Advanced Science and Technology, University of Tokyo, ⁹Institute for Advanced Biosciences, Keio University, ¹⁰Graduate School of Media and Governance, Keio University, ¹¹Department of Molecular Oncology, Graduate School of Medicine, Chiba University, ¹²Department of Biological Sciences, School of Science, University of Tokyo, ¹³College of Arts and Sciences, University of Tokyo

Here, we present a modified method for cryopreservation of one-cell embryos as well as a protocol that couples the use of freeze-thawed embryos and electroporation for the efficient generation of genetically modified mice.