RNA Detection and Visualization Methods and Protocols

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Of all biological molecules, RNA has perhaps the most complex and extensively regulated metabolism. Beginning with control at the transcriptional level, RNAs undergo nuclear processing, export, transport and localization, translation, storage, silencing and, ultimately, decay, all of which are mediated by cohorts of interacting proteins and additional RNA molecules. However, as most of the RNA life cycle actually occurs within the cytoplasm, it is imperative to have tools that allow for the localization of RNAs to be observed either in an in situ setting or, preferably, under in vivo conditions. In addition, there is a requisite need for tools that allow for both RNA purification from cells and the identification of bound interacting proteins and accessory RNAs. Finally, bioinformatic tools are requisite for mining the sequence and structural motifs necessary for RNAs to localize. When used in combination, these approaches can allow researchers to ask complex questions regarding where and how RNAs localize, and whether localization pertains to translational control, protein localization and complex assembly, and RNA stability. The aim of this volume is to provide an up-to-date and in-depth description of basic methods and protocols used for detecting and visualizing mRNAs in both fixed and live cells, from bacteria to mammals. Written for novices and experts alike, this mix of classic in situ hybridization and advanced live imaging techniques, cell fractionation and affinity purification procedures, and bioinformatic tools is to give researchers the most complete and extensive array of research aids possible. This volume is composed of twenty eight chapters separated into six subject areas: (1) Visualizing mRNAs in situ; (2) Visualizing mRNAs in vivo using molecular probes or reconstituted fluorescent proteins; (3) Visualizing mRNAs in vivo using aptamers and intact fluorescent proteins; (4) Use of cell fractionation to demonstrate the subcellular localization of RNA; (5) Affinity purification of mRNAs and the identification of trans-acting factors; and (6) Use of bioinformatics to identify cis-acting motifs and structures in RNAs. Chapters have been contributed by some of the best and brightest investigators, established or young, that have made an impact in the study of RNA localization and its importance in biological processes. Finally, I offer my sincere gratitude to the authors who have endeavored to make this book a unique and extensive collection of useful RNA detection methods and protocols for the benefit of other researchers.

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Informasi Detail

Judul Seri

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No. Panggil

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Penerbit

Kogakusha : Gramedia Pustaka Utama., 2011

Deskripsi Fisik

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Bahasa

English

ISBN/ISSN

978-1-61779-004-1

Klasifikasi

NONE

Tipe Isi

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Tipe Media

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Tipe Pembawa

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Edisi

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Subjek

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Info Detail Spesifik

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Pernyataan Tanggungjawab

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