Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
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.
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 Tranillegalscription 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 Wühr
14. Quantitative Proteomics for Xenopus Embryos II, Data Analysis
Matthew Sonnett, Meera Gupta, Thao Nguyen, and Martin Wühr
15. Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System
Lukas Weiss, Thomas Offner, Thomas Hassenklöver, and Ivan Manzini
16. X-FaCT: Xenopus-Fast Clearing Technique
Pierre Affaticati, Sébastien Le Mével, Arnim Jenett, Laurie Rivière, 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
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.
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.
Husbandry, General Care, and Transportation of Xenopus laevis, and Xenopus tropicalis.- Generation and Care of Xenopus laevis and Xenopus tropcalis Embryos.- Methods for CRISPR/Cas9 Xenopus tropicalis Tissue-Specific Multiplex Genome Engineering.- Targeted Genome Engineering in Xenopus using the Tranillegalscription activator-Like Effector Nuclease (TALEN) Technology.- Genotyping of CRISPR/Cas9 Genome Edited Xenopus tropicalis.- BATCH-GE: Analysis of NGS Data for Genome Editing Assessment.- A Simple Knock-In System for Xenopus via Microhomology-Mediated End Jointing Repair.- How to Generate Non-Mosaic CRSPR/Cas9 Mediated Knock-In and Mutations F0 Xenopus through the Host-Transfer Technique.- Targeted Electroporation in the CNS in Xenopus Embryos.- Conditional Chemogenetic Ablation of Photoreceptor Cells in Xenopus retina.- Cancer Models in Xenopus tropicalis by CRISPR/Cas 9 Mediated Knockout of Tumor Suppressors.- CRISPR/Cas9 F0 Screening of Congenital Heart Disease Genes in Xenopus tropicalis.- Quantitative Proteomics of Xenopus Embryos I, Sample Preparation.- Quantitative Proteomics for Xenopus Embryos II, Data Analysis.- Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System.- X-FaCT: Xenopus-Fast Clearing Technique.- Cell Cycle Analysis of the Embryonic Brain of Fluorescent Reporter Xenopus tropicalis by Flow Cytometry.- Manipulating and Analyzing Cell Type Composition of the Xenopus Mucociliary Epidermis.- Evaluating Blood Cell Populations in Xenopus using Flow Cytometry and Differential Counts by Cytospin.- Isolation and Culture of Amphibian (Xenopus laevis) Sub-Capsular Liver and Bone Marrow Cells.- Isolation and Primary Culture Methods of Adult and Larval Myogenic Cells from Xenopus laevis.
Inhaltsverzeichnis
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 Tranillegalscription 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 Wühr
14. Quantitative Proteomics for Xenopus Embryos II, Data Analysis
Matthew Sonnett, Meera Gupta, Thao Nguyen, and Martin Wühr
15. Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System
Lukas Weiss, Thomas Offner, Thomas Hassenklöver, and Ivan Manzini
16. X-FaCT: Xenopus-Fast Clearing Technique
Pierre Affaticati, Sébastien Le Mével, Arnim Jenett, Laurie Rivière, 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
Klappentext
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.rnrnCutting-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.
rn
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
