Transgenesis: Methods and Protocols

ISBN-13: 9781071629895 / Angielski / Twarda / 2023 / 437 str.

Thomas L. Saunders

Opis

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.

Spis treści

1. Transgenesis and Genome Engineering: A Historical Review

            Lluis Montoliu

 

2. Practical Application of the 3Rs in Rodent Transgenesis

            Thorsten Buch, Boris Jerchow, and Branko Zevnik

 

3. Genetic and Molecular Quality Control of Genetically Engineered Mice

            Lauri Lintott and Lauryl M.J. Nutter

 

4. Genotyping Genome-Edited Founders and Subsequent Generation

            Matthew Mackenzie, Alex Fower, Alasdair J. Allan, Gemma F. Codner, Rosie K. Bunton-Stasyshyn, and Lydia Teboul

 

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

            Charles Yu, Roger Caothien, Anna Pham, Lucinda Tam, Tuija Alcantar, Natasha Bacarro, Juan Reyes Jr., Marques Jackson, Brian Nakao, and Merone Roose-Girma

 

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

            Shilpa Narina, Jon P. Connelly, and Shondra M. Pruett-Miller

 

7. Advanced Technologies and Automation in mES Cell Workflow

            Charles Yu, Roger Caothien, Marques Jackson, Brian Nakao, Anna Pham, Lucinda Tam, and Merone Roose-Girma

 

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

            Benedikt Wefers, Wolfgang Wurst, and Ralf Kühn

 

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

            Mia Wallace, J. Michael White, Evgenea Kouranova, Zi Teng Wang, and Xiaoxia Cui

 

10. CRISPR/Cas9 Endonuclease Mediated Mouse Genome Editing of 1 Cell and/or 2 Cell Embryos by Electroporation, and the Use of Rad51 to Enhance Knock-In Allele Homozygosity via Inter Homolog Repair Mechanism

            Raehum Paik and Selika Garza

 

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

            Daniel J. Davis, Hongsheng Men, and Elizabeth C. Bryda

 

12. CRISMERE Chromosome Engineering in Mouse and Rat

            Laurence Schaeffer, Loic Lindner, Guillaume Pavlovic, Yann Hérault, and Marie-Christine Birling

 

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

            Daniel Biggs, Chiann-mun Chen, and Benjamin Davies

 

14. 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, Shigeru Kakuta, and Masato Ohtsuka

 

15. Gene Targeting in Rat Embryonic Stem Cells

            Hongsheng Men, Daniel J. Davis, and Elizabeth C. Bryda

 

16. Rat Embryonic Stem Cell Transgenesis

            Elizabeth C. Bryda, Hongsheng Men and Barbara J. Stone

 

17. ­CRISPR/Cas9-Mediated Genome Editing in Zebrafish

            Jeffrey G. Daniel, Xinge Yu, Allison C. Ferguson, and Jordan A. Shavit

 

18. Generation of Rabbit Chimeras by 8-Cell Stage Embryo Injection

            Dongshan Yang, Jun Song, Jie Xu, Jifeng Zhang, and Y. Eugene Chen

 

19. Genome Editing in Pigs

            David Preisinger, Thomas Winogrodzki, Bernhard Klinger, Angelika Schnieke, and Beate Rieblinger

 

20. Generation of Genome-Edited Chicken through Targeting of Primordial Germ Cells

            Alewo Idoko-Akoh and Michael J. McGrew