Synopses & Reviews
It is now known that microRNA (miRNA), once thought to be junk, can suppress the expression of other genes and may be involved in numerous cellular processes, including cell proliferation, apoptosis, cell differentiation, and development. In MicroRNA Protocols, expert miRNA researchers explain its basic concepts and introduce the most advanced technologies and techniques for predicting screening, isolating, and assaying it. The authors provide diverse, novel, and useful descriptions of miRNAs in several species-including plants, worms, flies, fish, chicks, mice, and humans-and show how they have been employed to develop miRNA as a potential drug design tool. Highlights include miRNA and their target prediction by computer analysis, functional assays of miRNA in vitro and in vivo, and transgenic animal models using miRNA technologies. Additional chapters cover gene regulation, small RNA function, RNA interference mechanisms, and transgenetics. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Eminently practical and cutting-edge, MicroRNA Protocols affords biomedical researchers readily reproducible techniques to understand and study the molecular pathogenesis of disease and design new therapeutic strategies.
Table of Contents
The MicroRNA: Overview of the RNA Gene That Modulates Gene Functions Shao-Yao Ying, Donald C. Chang, Joseph D. Miller, and Shi-Lung Lin Structure Analysis of MicroRNA Precursors Jacek Krol and Wlodzimierz J. Krzyzosiak MicroRNA Biogenesis: Isolation and Characterization of the Microprocessor Complex Richard I. Gregory, Thimmaiah P. Chendrimada, and Ramin Shiekhattar Recognition and Cleavage of Primary MicroRNA Transcripts Yan Zeng and Bryan R. Cullen Mouse Embryonic Stem Cells as a Model Genetic System to Dissect and Exploit the RNA Interference Machinery Stefan A. Muljo and Chryssa Kanellopoulou MicroRNAs and Messenger RNA Turnover Julia A. Chekanova and Dmitry A. Belostotsky Prediction of MicroRNA Targets Marc Rehmsmeier Prediction of Human MicroRNA Targets Bino John, Chris Sander, and Debora S. Marks Complications in Mammalian MicroRNA Target Prediction Neil R. Smalheiser and Vetle I. Torvik miRBase:The MicroRNA Sequence Database Sam Griffiths-Jones Methodologies for High-Throughput Expression Profiling of MicroRNAs Paz Einat In Situ Hybridization as a Tool to Study the Role of MicroRNAs in Plant Development Catherine Kidner and Marja Timmermans Usefulness of the Luciferase Reporter System to Test the Efficacy of siRNA Fengfeng Zhuang and Yi-Hsin Liu Cloning MicroRNAs From Mammalian Tissues Michael Z. Michael Methods for Analyzing MicroRNA Expression and Function During Hematopoietic Lineage Differentiation Hyeyoung Min and Chang-Zheng Chen Identifying MicroRNA Regulators of Cell Death in Drosophila Chun-Hong Chen, Ming Guo, and Bruce A. Hay MicroRNAs in Human Immunodeficiency Virus-1 Infection Yamina Bennasser, Shu-Yun Le, Man Lung Yeung, and Kuan-Teh Jeang Cloning and Detection of HIV-1-Encoded MicroRNA Shinya Omoto and Yoichi R. Fujii Identification of Messenger RNAs and MicroRNAs Associated With Fragile X Mental Retardation Protein Ranhui Duan and Peng Jin In Vitro Precursor MicroRNA Processing Assays Using Drosophila Schneider-2 Cell Lysates Akira Ishizuka, Kuniaki Saito, Mikiko C. Siomi, and Haruhiko Siomi Downregulation of Human Cdc6 Protein Using a Lentivirus RNA Interference Expression Vector Feng Luo, Jiing-Kuan Yee, Sheng-He Huang, Ling-Tao Wu, and Ambrose Y. Jong Gene Silencing In Vitro and In Vivo Using Intronic MicroRNAs Shi-Lung Lin and Shao-Yao Ying Isolation and Identification of Gene-Specific MicroRNAs Shi-Lung Lin, Donald C. Chang, and Shao-Yao Ying Transgene-Like Animal Models Using Intronic MicroRNAs Shi-Lung Lin, Shin-Ju E. Chang, and Shao-Yao Ying Evolution of MicroRNAs Andrea Tanzer and Peter F. Stadler Perspectives Shao-Yao Ying, Donald C. Chang, Joseph D. Miller, and Shi-Lung Lin Index