Table Of ContentXenopus tropicalis as a Model Organism for Genetics and Genomics: Past, Present and Future.-Husbandry of Xenopus tropicalis .-Generating Diploid Embryos from Xenopus tropicalis .-Navigating the Xenopus tropicalis Genome.-Genetic Analysis of Xenopus tropicalis .-Forward Genetic Screens in Xenopus using Transposon-mediated Insertional Mutagenesis.-Targeted Gene Disruption with Engineered Zinc-finger Nucleases (ZFNs).-Reverse Genetic Studies using Antisense Morpholino Oligonucleotides.-Chemical Genetics and Drug Discovery in Xenopus.-Maternal mRNA Knock-down Studies: Antisense Experiments using the Host-transfer Technique in Xenopus laevis and Xenopus tropicalis .-Generating Transgenic Frog Embryos by Restriction Enzyme Mediated Integration (REMI).-A Simple Method of Transgenesis using I- Sce I meganuclease in Xenopus .-Using FC31 Integrase to Mediate Insertion of DNA in Xenopus Embryos.- Xenopus Transgenics: Methods using Transposons.-Comparative Genomics-based Identification and Analysis of Cis-regulatory Elements.-Tet-On Binary Systems for Tissue-specific and Inducible Transgene Expression.-Chromatin Immunoprecipitation Analysis of Xenopus Embryos.-Transcriptomics Using Next Generation Sequencing Technologies.-Databases of Gene Expression in Xenopus Development.-Investigating Alternative RNA Splicing in Xenopus .-Immunoisolation of Protein Complexes from Xenopus .-Complementary Proteomic Analysis of Protein Complexes.-Antibody Development and use in Chromogenic and Fluorescent Immunostaining.-Multiple fluorescent in situ mRNA Hybridization (FISH) on Whole Mounts and Sections.-Methods to Analyze microRNA Expression and Function During Xenopus Development.-A Bromodeoxyuridine (BrdU) Based Protocol for Characterizing Proliferating Progenitors in Xenopus Embryos.-Microscopy Tools for Quantifying Developmental Dynamics in XenopusEmbryos.-Mathematical Modelling of Gene RegulatoryNetworks in Xenopus Development.-Stem-cell-like Embryonic Explants to Study Cardiac Development.-Studying Regeneration in Xenopus .-On-line Resources for Xenopus. Mathematical Modelling of Gene RegulatoryNetworks in Xenopus Development.-Stem-cell-like Embryonic Explants to Study Cardiac Development.-Studying Regeneration in Xenopus .-On-line Resources for Xenopus.
SynopsisThe 2nd edition of this Methods in Molecular Biology(tm) volume provides novel approaches inspired by Xenopus tropicalis and the sequencing of its genome. Offers a toolkit enabling laboratories to maximize the power of this extraordinary experimental system., The 2nd edition of this Methods in Molecular Biology(t) volume provides novel approaches inspired by Xenopus tropicalis and the sequencing of its genome. Offers a toolkit enabling laboratories to maximize the power of this extraordinary experimental system., Part I: Xenopus Tropicalis 1. Xenopus tropicalis as a Model Organism for Genetics and Genomics: Past, Present and Future Robert M. Grainger 2. Husbandry of Xenopus tropicalis Alan Jafkins, Anita Abu-Daya, Anna Noble, Lyle Zimmerman, and Matthew Guille 3. Generating Diploid Embryos from Xenopus tropicalis Florencia del Viso and Mustafa Khokha 4. Navigating the Xenopus tropicalis Genome Ira L. Blitz Part II: Genetics in Xenopus 5. Genetic Analysis of Xenopus tropicalis Timothy J. Geach, Derek L. Stemple, and Lyle B. Zimmerman 6. Forward Genetic Screens in Xenopus using Transposon-mediated Insertional Mutagenesis Donald A. Yergeau, Clair M. Kelley, Haiqing Zhu, Emin Kuliyev and Paul E. Mead 7. Targeted Gene Disruption with Engineered Zinc-finger Nucleases (ZFNs) John J. Young and Richard M. Harland 8. Reverse Genetic Studies using Antisense Morpholino Oligonucleotides Yanan Zhao, Shoko Ishibashi and Enrique Amaya 9. Chemical Genetics and Drug Discovery in Xenopus Matthew L. Tomlinson, Adam Hendry, and Grant N. Wheeler 10. Maternal mRNA Knock-down Studies: Antisense Experiments using the Host-transfer Technique in Xenopus laevis and Xenopus tropicalis David J. Olson, Alissa M. Hulstrand and Douglas W. Houston Part II: Xenopus Trangenesis 11. Generating Transgenic Frog Embryos by Restriction Enzyme Mediated Integration (REMI) Shoko Ishibashi, Kristen L. Kroll and Enrique Amaya 12. A Simple Method of Transgenesis using I-SceI meganuclease in Xenopus Shoko Ishibashi, Nick R. Love and Enrique Amaya 13. Using FC31 Integrase to Mediate Insertion of DNA in Xenopus Embryos You E. Li, Bryan G. Allen and Daniel L. Weeks 14. Xenopus Transgenics: Methods using Transposons Clair M. Kelley, Donald A. Yergeau, Haiqing Zhu, Emin Kuliyev and Paul E. Mead 15. Comparative Genomics-based Identification and Analysis of Cis-regulatory Elements Hajime Ogino, Haruki Ochi, Chihiro Uchiyama, Sarah Louie, and Robert M. Grainger 16. Tet-On Binary Systems for Tissue-specific and Inducible Transgene Expression Daniel R. Buchholz Part III: Gene Expression: From Gene to Transcript to Protein 17. Chromatin Immunoprecipitation Analysis of Xenopus Embryos Robert C. Akkers, Ulrike G. Jacobi and Gert Jan C. Veenstra 18. Transcriptomics Using Next Generation Sequencing Technologies Dasfne Lee-Liu, Leonardo I. Almonacid, Fernando Faune, Francisco Melo, Juan Larrain 19. Databases of Gene Expression in Xenopus Development Michael J. Gilchrist, Nicolas Pollet 20. Investigating Alternative RNA Splicing in Xenopus Agnès Mereau and Serge Hardy 21. Immunoisolation of Protein Complexes from Xenopus Frank L. Conlon, Yana Miteva, Erin Kaltenbrun, Lauren Waldron, Todd M. Greco, Ileana M. Cristea 22. Complementary Proteomic Analysis of Protein Complexes Todd M. Greco, Yana Miteva, Frank L. Conlon, Ileana M. Cristea Part IV: Imaging Xenopus Development 23. Antibody Development and use in Chromogenic and Fluorescent Immunostaining Eamon Dubaissi, Niki Panagiotaki, Nancy Papalopulu, and Peter D. Vize 24. Multiple fluorescent in situ mRNA Hybridization (FISH) on Whole Mounts and Sections Robert Lea, Boyan Bonev, Eamon Dubaissi, Peter D. Vize and Nancy Papalopulu 25. Methods to Analyze microRNA Expression and Function During Xenopus Development Boyan Bonev and Nancy Papalopulu 26. A Bromodeoxyur, During the 20th century Xenopus has flourished as an accessible experimental model system for studying vertebrate biology. The second edition of Xenopus Protocols:Post-Genomic Approaches expands upon the previous edition with novel approaches inspired by Xenopus tropicalis and the sequencing of its genome . Xenopus tropicalis genomics and tractable genetics combined with the unique experimental advantages of Xenopus laevis and their distinctive phylogenetic position make them unique model systems for vertebrate embryonic development in the 21st century. Written in the highly successful Methods in Molecular Biology(tm) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Xenopus Protocols: Post-Genomic Approaches, Second Edition provides a toolkit that will enable every laboratory to maximize the power of this extraordinary experimental system in the post-genomic age.