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Cairo University

4 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.

Chemistry

Synthesis of Black Seed Oil-loaded-, Alginate-based, pH-sensitive Beads Using Electrospraying Technique

Abul Kalam Azad¹, Mohamad Dayoob², Vinoth Kumarasamy³, Vetriselvan Subramaniyan⁴, Ghadeer M. Albadrani⁵, Muath Q. Al-Ghadi⁶, Mohamed Kamel⁷, Mohamed M. Abdel-Daim^8,9, Mireia Mallandrich Miret¹⁰, Abd Almonem Doolaanea^11,12

¹Faculty of Pharmacy, University College of MAIWP International, ²Faculty of Pharmacy, MAHSA University, ³Department of Parasitology and Medical Entomology, Faculty of Medicine, Universiti Kebangsaan Malaysia, ⁴Department of Medical Sciences, School of Medical and Life Sciences, Sunway University, ⁵Department of Biology, College of Science, Princess Nourah bint Abdulrahman University, ⁶Department of Zoology, College of Science, King Saud University, ⁷Department of Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, ⁸Department of Pharmaceutical Sciences, Pharmacy Program, Batterjee Medical College, ⁹Pharmacology Department, Faculty of Veterinary Medicine, Suez Canal University, ¹⁰Department of Pharmacy, Pharmaceutical Technology and Physical-Chemistry, Faculty of Pharmacy and Food Sciences, University of Barcelona, ¹¹Research and Development, Sabrena Experience, ¹²Alphastar Lab Systems

A technique employing high electric voltage and a targeted, active ingredient-loaded emulsion to fabricate pH-responsive, uniform microbeads is presented.