Main description:

This volume explores techniques used to study and experiment with Xenopus in order to model or understand human disease. The chapters in this book cover topics such as implementation of CRISPR/Cas9 and TALEN in Xenopus from the design stage up to the genotyping stage; methods to explore generation of knock-in animals; cancer modelling and in vivo screening of congenital heart disease; electroporation; and phenotyping at the organismal level, cellular level, and proteome level. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Xenopus: Methods and Protocols is a valuable resource for researchers who are interested in expanding their knowledge of this wide and developing field. This book will aid in keeping Xenopus at the forefront for biochemical, cell biological, and developmental studies and forward it as a preclinical disease model.

Contents:

Preface...

Table of Contents...

Contributing Authors...

1. Husbandry, General Care, and Transportation of Xenopus laevis, and Xenopus tropicalis

Sean McNamara, Marcin Wlizla, and Marko E. Horb

2. Generation and Care of Xenopus laevis and Xenopus tropcalis Embryos

Marcin Wlizla, Sean McNamara, and Marko E. Horb

3. Methods for CRISPR/Cas9 Xenopus tropicalis Tissue-Specific Multiplex Genome Engineering

Thomas Naert and Kris Vleminckx

4. Targeted Genome Engineering in Xenopus using the Transcription activator-Like Effector Nuclease (TALEN) Technology

Tom Van Nieuwenhuysen and Kris Vleminckx

5. Genotyping of CRISPR/Cas9 Genome Edited Xenopus tropicalis

Thomas Naert and Kris Vleminckx

6. BATCH-GE: Analysis of NGS Data for Genome Editing Assessment

Wouter Steyaert, Annekatrien Boel, Paul Coucke, and Andy Willaert

7. A Simple Knock-In System for Xenopus via Microhomology-Mediated End Jointing Repair

Ken-ich T. Suzuki, Yuto Sakane, Miyuki Suzuki, and Takashi Yamamoto

8. How to Generate Non-Mosaic CRISPR/Cas9 Mediated Knock-In and Mutations F0 Xenopus through the Host-Transfer Technique

Emmanuel Tadjuidje and Sang-Wook Cha

9. Targeted Electroporation in the CNS in Xenopus Embryos

Hovy Ho-Wai Wong and Christine E. Holt

10. Conditional Chemogenetic Ablation of Photoreceptor Cells in Xenopus retina

Albert Chesneau, Odile Bronchain, and Muriel Perron

11. Cancer Models in Xenopus tropicalis by CRISPR/Cas 9 Mediated Knockout of Tumor Suppressors

Thomas Naert and Kris Vleminckx

12. CRISPR/Cas9 F0 Screening of Congenital Heart Disease Genes in Xenopus tropicalis

Engin Deniz, Emily K. Mis, Muara Lane, and Mustafa K. Khokha

13. Quantitative Proteomics of Xenopus Embryos I, Sample Preparation

Meera Gupta, Matthew Sonnett, Lillia Ryazanova, Marc Presler, and Martin Wuhr

14. Quantitative Proteomics for Xenopus Embryos II, Data Analysis

Matthew Sonnett, Meera Gupta, Thao Nguyen, and Martin Wuhr

15. Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System

Lukas Weiss, Thomas Offner, Thomas Hassenkloever, and Ivan Manzini

16. X-FaCT: Xenopus-Fast Clearing Technique

Pierre Affaticati, Sebastien Le Mevel, Arnim Jenett, Laurie Riviere, Elodie Machado, Bilal B. Mughal, and Jean-Baptiste Fini

17. Cell Cycle Analysis of the Embryonic Brain of Fluorescent Reporter Xenopus tropicalis by Flow Cytometry

Rivka Noelanders and Kris Vleminckx

18. Manipulating and Analyzing Cell Type Composition of the Xenopus Mucociliary Epidermis

Peter Walentek

19. Evaluating Blood Cell Populations in Xenopus using Flow Cytometry and Differential Counts by Cytospin

Jacques Robert, Eva-Stina Edholm, and Francisco De Jesus Andino

20. Isolation and Culture of Amphibian (Xenopus laevis) Sub-Capsular Liver and Bone Marrow Cells

Amulya Yaparla and Leon Grayfer

21. Isolation and Primary Culture Methods of Adult and Larval Myogenic Cells from Xenopus laevis

Kazi Taheruzzaman and Akio Nishikawa