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Transgenesis

Methods and Protocols

  • Book
  • © 2023

Overview

Editors:
  1. Thomas L. Saunders

    1. Transgenic Animal Model Core, University of Michigan, Ann Arbor, USA

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  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2631)

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Table of contents (21 protocols)

  1. Front Matter

    Pages i-xii
    Download chapter PDF

  2. Transgenesis and Genome Engineering: A Historical Review

    • Lluis Montoliu
    Pages 1-32

  3. Practical Application of the 3Rs in Rodent Transgenesis

    • Thorsten Buch, Boris Jerchow, Branko Zevnik
    Pages 33-51

  4. Genetic and Molecular Quality Control of Genetically Engineered Mice

    • Lauri G. Lintott, Lauryl M. J. Nutter
    Pages 53-101

  5. Genotyping Genome-Edited Founders and Subsequent Generation

    • Matthew Mackenzie, Alex Fower, Alasdair J. Allan, Gemma F. Codner, Rosie K. Bunton-Stasyshyn, Lydia Teboul
    Pages 103-134

  6. ASIS-Seq: Transgene Insertion Site Mapping by Nanopore Adaptive Sampling

    • Charles Yu, Roger Caothien, Anna Pham, Lucinda Tam, Tuija Alcantar, Natasha Bacarro et al.
    Pages 135-153

  7. High-Throughput Analysis of CRISPR-Cas9 Editing Outcomes in Cell and Animal Models Using CRIS.py

    • Shilpa Narina, Jon P. Connelly, Shondra M. Pruett-Miller
    Pages 155-182

  8. Advanced Technologies and Automation in mES Cell Workflow

    • Anna Pham, Roger Caothien, Lucinda Tam, Diego Diaz, Marques Jackson, Brian Nakao et al.
    Pages 183-206

  9. Gene Editing in Mouse Zygotes Using the CRISPR/Cas9 System

    • Benedikt Wefers, Wolfgang Wurst, Ralf Kühn
    Pages 207-230

  10. Floxing by Electroporating Single-Cell Embryos with Two CRISPR RNPs and Two ssODNs

    • Mia Wallace, J. Michael White, Evgenea Kouranova, Zi Teng Wang, Xiaoxia Cui
    Pages 231-252

  11. CRISPR/Cas9 Endonuclease-Mediated Mouse Genome Editing of One-Cell and/or Two-Cell Embryos by Electroporation, and the Use of Rad51 to Enhance Knock-In Allele Homozygosity via Interhomolog Repair Mechanism

    • Selika Garza, Raehum Paik
    Pages 253-266

  12. Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes

    • Daniel J. Davis, Hongsheng Men, Elizabeth C. Bryda
    Pages 267-276

  13. CRISMERE Chromosome Engineering in Mouse and Rat

    • Laurence Schaeffer, Loic Lindner, Guillaume Pavlovic, Yann Hérault, Marie-Christine Birling
    Pages 277-297

  14. Targeted Integration of Transgenes at the Mouse Gt(ROSA)26Sor Locus

    • Daniel Biggs, Chiann-mun Chen, Benjamin Davies
    Pages 299-323

  15. Improved Genome Editing via Oviductal Nucleic Acids Delivery (i-GONAD): Protocol Steps and Additional Notes

    • Masahiro Sato, Ayaka Nakamura, Marie Sekiguchi, Takashi Matsuwaki, Hiromi Miura, Channabasavaiah B. Gurumurthy et al.
    Pages 325-340

  16. Gene Targeting in Rat Embryonic Stem Cells

    • Hongsheng Men, Daniel J. Davis, Elizabeth C. Bryda
    Pages 341-353

  17. Rat Embryonic Stem Cell Transgenesis

    • Elizabeth C. Bryda, Hongsheng Men, Barbara J. Stone
    Pages 355-370

  18. CRISPR/Cas9-Mediated Genome Editing in Zebrafish

    • Jeffrey G. Daniel, Xinge Yu, Allison C. Ferguson, Jordan A. Shavit
    Pages 371-380

  19. Generation of Rabbit Chimeras by Eight-Cell Stage Embryo Injection

    • Dongshan Yang, Jun Song, Jie Xu, Jifeng Zhang, Y. Eugene Chen
    Pages 381-391

  20. Genome Editing in Pigs

    • David Preisinger, Thomas Winogrodzki, Bernhard Klinger, Angelika Schnieke, Beate Rieblinger
    Pages 393-417
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Keywords

About this book

This detailed volume focuses on genotyping and validation in addition to information on how to produce gene edited cells and animals for research. Future advances in biomedical research will benefit greatly from the use of precise gene targeting of transgenes in the genome as CRISPR technology supersedes earlier methods that relied on random transgene integration, which this collection reflects. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips for troubleshooting and avoiding known pitfalls. 

Authoritative and practical, Transgenesis: Methods and Protocols serves as an ideal guide for researchers working with or working on genetically modified models.

Editors and Affiliations

  • Transgenic Animal Model Core, University of Michigan, Ann Arbor, USA

    Thomas L. Saunders

Bibliographic Information

  • Book Title: Transgenesis

  • Book Subtitle: Methods and Protocols

  • Editors: Thomas L. Saunders

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-2990-1

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2023

  • Hardcover ISBN: 978-1-0716-2989-5Published: 31 March 2023

  • Softcover ISBN: 978-1-0716-2992-5Published: 31 March 2023

  • eBook ISBN: 978-1-0716-2990-1Published: 30 March 2023

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XII, 448

  • Number of Illustrations: 17 b/w illustrations, 104 illustrations in colour

  • Topics: Biological Techniques, Genetics and Genomics

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