Protocol


Part of the
Methods in Molecular Biology
book series (MIMB, volume 2312)

Abstract

Developments in genome-editing technology, especially CRISPR-Cas9, have revolutionized the way in which genetically engineered animals are generated. However, the process of generation includes microinjection to the one-cell stage embryo and the transfer of the microinjected embryo to the surrogate animals, which requires trained personnel. We recently reported the method includes introduction of CRISPR-Cas9 systems to the developing cerebral cortex via in utero electroporation thus generating gene-targeted neural stem cells in vivo. This technique is widely applicable for gene knockout, monitoring gene expression, and lineage analysis in developmental biology. In this chapter, the detailed protocol of EGFP (enhanced green fluorescent protein) knock-in method via in utero electroporation is described.

Key words

De novo gene targeting In utero electroporation CRISPR-Cas9 Developmental biology Visualization of protein subcellular localization 

Supplementary material

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Laboratory for Cell AsymmetryRIKEN Center for Biosystems Dynamics ResearchKobeJapan
  2. 2.Division of Molecular and Medical Genetics, Institute of Medical ScienceUniversity of TokyoTokyoJapan
  3. 3.Public Relations Division, Office of Global CommunicationsKyoto UniversityKyotoJapan
  4. 4.Laboratory of Molecular Cell Biology and Development, Department of Animal Development and Physiology, Graduate School of BiostudiesKyoto UniversityKyotoJapan



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