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

9.6K Views

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06:46 min

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April 2nd, 2020

DOI :

10.3791/60808-v

April 2nd, 2020

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Hirofumi Nishizono*¹^,²^,³, Mohamed Darwish*⁴^,⁵, Hideki Uosaki⁶^,⁷, Nanami Masuyama⁸^,⁹^,¹⁰, Motoaki Seki⁸^,¹¹, Hiroyuki Abe³, Nozomu Yachie⁸^,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

Summary

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Freeze thawed Embryos

Genetically Modified Mice

High efficiency Production

Electroporation

C57BL 6J Mice

In Vitro Fertilization

Super Ovulation

Human Chorionic Gonadotropin

Capacitation

Cumulus Oocyte Complexes

Chapters in this video

0:05

Introduction

0:35

In Vitro Fertilization

2:23

Embryo Cryopreservation

3:13

Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) Reagent Assembly

4:19

CRISPR Electroporation

5:09

Results: Representative Knockout Mouse Generation Efficiency and Mosaic Rate Evaluation

6:09

Conclusion

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