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As the number of sequenced genomes continues to increase, understanding the functions of newly discovered molecules will require greater efficiency and further study within the context of live cells. In Microinjection: Methods and Applications, expert researchers contribute methods utilizing microinjection techniques ranging from expression of RNA to the integration of DNA into the genome with the ultimate goal of learning about gene expression, signal transduction, and protein function within these living cells. This versatile volume updates established techniques such as cRNA expression in Xenopus oocytes, and examines new, cutting-edge technologies, including antisense morpholino oligonucleotides, RNAi for knockdown experiments, and the use of integrase to produce transgenic animals, all through microinjection techniques that can be easily adopted by any lab. Written in the highly successful Methods in Molecular Biology'„¢ series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, highlighting tips on troubleshooting and avoiding known pitfalls. Comprehensive and easily accessible, Microinjection: Methods and Applications is an ideal source for new ideas and techniques certain to aid in the advance of biological research.
Table of Contents
1. Expression of Exogenous mRNA in Xenopus laevis Embryos for the Study of Cell Cycle RegulationJill C. Sible and Brian N. Wroble2. Use of Luciferase Chimaera to Monitor PLC Expression in Mouse EggsKarl Swann, Karen Campbell, Yuansong Yu, Christopher Saunders, and F. Anthony Lai3. Analysis of 14-3-3 Family Member Function in Xenopus Embryos by Microinjection of Antisense Morpholino OligosJeffrey M. C. Lau and Anthony J. Muslin4. A Microinjectable Biological System, the Xenopus Oocyte, as an Approach to Understanding Signal Transduction Protein FunctionKatia Cailliau and Edith Browaeys-Poly5. Combining Microinjection and Immunoblotting to Analyze MAP Kinase Phosphorylation in Single Starfish Oocytes and Eggs David J. Carroll and Wei Hua6. Analysis of Signaling Pathways in Zebrafish Development by MicroinjectionWilliam H. Kinsey7. Protein Inhibition by Microinjection and RNA-Mediated Interference in Tissue Culture Cells: Complimentary Approaches to Study Protein FunctionJane R. Stout, Rania S. Rizk, and Claire E. Walczak8. DNA Delivery by Microinjection for the Generation of Recombinant Mammalian Cell LinesSebastien Chenuet, Madiha Derouazi, David Hacker, and Florian Wurm9. Bacteriophage fC31 Integrase Mediated Transgenesis in Xenopus laevis for Protein Expression at Endogenous LevelsBryan G. Allen and Daniel L. Weeks10. Germline Transformation of Caenorhabditis elegans by InjectionPavan Kadandale, Indrani Chatterjee, and Andrew Singson11. Quantitative Microinjection of Mouse Oocytes and EggsDouglas Kline12. Microinjection of Follicle-Enclosed Mouse OocytesLaurinda A. Jaffe, Rachael P. Norris, Marina Freudzon, William J. Ratzan, and Lisa M. Mehlmann13. Functional Studies of Regulatory Genes in the Sea Urchin EmbryoVincenzo Cavalieri, Maria Di Bernardo, and Giovanni Spinelli14. Exploring the Cytoskeleton during Intracytoplasmic Sperm Injection in HumansVanesa Y. Rawe and Héctor Chemes15. Somatic Cell Nuclear Transfer in the MouseSatoshi Kishigami and Teruhiko Wakayama