In this book 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.

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.

Explores a wide variety of model systems, including human, mouse, sea urchin, starfish, zebrafish, and Xenopus oocytes, transformed Caenorhabditis elegans, and tissue culture cells Contains detailed methods easily adaptable to virtually any system Covers gene and protein function in living cells, analysis of signal transduction pathways in vivo, single cell analysis, systems biology in a living cell, quantitative analysis of signaling transduction, antisense morpholino oligonucleotide and RNAi analysis, quantitative microinjection, and analysis of Gene Expression, among other topics Expands the capabilities of cellular and molecular biology laboratories and provides scientists with a valuable mechanistic understanding Includes supplementary material: sn.pub/extras

Inhalt
Expression of Exogenous mRNA in Xenopus laevis Embryos for the Study of Cell Cycle Regulation.- Use of Luciferase Chimaera to Monitor PLC? Expression in Mouse Eggs.- Analysis of 14-3-3 Family Member Function in Xenopus Embryos by Microinjection of Antisense Morpholino Oligos.- A Microinjectable Biological System, the Xenopus Oocyte, as an Approach to Understanding Signal Transduction Protein Function.- Combining Microinjection and Immunoblotting to Analyze MAP Kinase Phosphorylation in Single Starfish Oocytes and Eggs.- Analysis of Signaling Pathways in Zebrafish Development by Microinjection.- Protein Inhibition by Microinjection and RNA-Mediated Interference in Tissue Culture Cells: Complementary Approaches to Study Protein Function.- DNA Delivery by Microinjection for the Generation of Recombinant Mammalian Cell Lines.- Bacteriophage ?C31 Integrase Mediated Transgenesis in Xenopus laevis for Protein Expression at Endogenous Levels.- Germline Transformation of Caenorhabditis elegans by Injection.- Quantitative Microinjection of Mouse Oocytes and Eggs.- Microinjection of Follicle-Enclosed Mouse Oocytes.- Functional Studies of Regulatory Genes in the Sea Urchin Embryo.- Exploring the Cytoskeleton During Intracytoplasmic Sperm Injection in Humans.- Somatic Cell Nuclear Transfer in the Mouse.