MRNA processing and metabolism : methods and protocols / edited by Daniel R. Schoenberg.

Contributor(s): Schoenberg, Daniel R
Material type: TextTextSeries: Methods in molecular biology (Clifton, N.J.): v. 257.Publisher: Totowa, N.J. : Humana Press, ©2004Description: 1 online resource (xv, 270 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 9781592597505; 1588292258; 9781588292254; 1592597505Subject(s): Messenger RNA -- Laboratory manuals | RNA, Messenger -- metabolism | Genetic Techniques | SCIENCE -- Life Sciences -- Molecular Biology | Messenger RNAGenre/Form: Electronic books. | Laboratory manuals. Additional physical formats: MRNA processing and metabolismDDC classification: 572.8/8 LOC classification: QP623.5.M47 | M765 2004Online resources: Click here to access online
Contents:
Using chromatin immunoprecipitation to map cotranscriptional mRNA processing in saccharomyces cerevisiae / Michael-Christopher Keogh and Stephen Buratowski -- Identifying phosphoCTD-associating proteins / Hemali P. Phatnani and Arno L. Greenleaf -- Imaging alternative splicing in living cells / Eric J. Wagner [and others] -- Tobramycin affinity tag purification of spliceosomes / Klaus Hartmuth, Hans-Peter Vornlocher and Reinhard Lührmann -- Using single-strand conformational polymorphism gel electrophoresis to analyze mutually exclusive alternative splicing / Alicia M. Celotto and Brenton R. Graveley -- Methods for the analysis of adenosine-to-inosine editing in RNA / Zuo Zhang and Gordon G. Carmichael -- Assaying nuclear messenger RNA export in human cells / Bryan R. Cullen -- RNA unwinding assay for DExD/H-Box RNA helicases / Stephanie S. Tseng-Rogenski and Tien-Hsien Chang -- Approaches for monitoring nuclear translation / Francisco J. Iborra, Dean A. Jackson, and Peter R. Cook -- Immunopurification and analysis of protein and RNA components of mRNP in mammalian cells / Fabrice Lejeune and Lynne E. Maquat.
Gene expression analysis of messenger RNP complexes / Luiz O.F. Penalva, Scott A. Tenenbaum, and Jack D. Keene -- Using the -N peptide to tether proteins to RNAs / Julie Baron-Benhamou [and others] -- An efficient system for cap- and poly(A)-dependent translation in vitro / Yuri V. Svitkin and Nahum Sonenberg -- A poly(A) tail-responsive in vitro system for cap- or IRES-driven translation from HeLa cells / Christian Thoma, Antje Ostareck-Lederer and Matthias W. Hentze -- Assessing messenger RNA decapping in cellular extracts / Naomi Bergman [and others] -- Analysis of RNA exonucleolytic activities in cellular extracts / Devi Mukherjee [and others] -- Application of ligation-mediated reverse transcription polymerase chain reaction to the identification of in vivo endonuclease-generated messenger RNA decay intermediates / Mark N. Hanson and Daniel R. Schoenberg -- Biochemical dissection of RNA silencing in plants / Guiliang Tang and Phillip D. Zamore -- RNA interference of mRNA processing factors in drosophila S2 cells / Alicia M. Celotto and Brenton R. Graveley -- RNA interference by short hairpin RNAs expressed in vertebrate cells / Gregory J. Hannon and Douglas S. Conklin.
Summary: MRNA processing is a key step in gene expression that effects all the proteins within the cell. In mRNA Processing and Metabolism: Methods and Protocols, world-renowned researchers bring together the latest techniques spanning the breadth of mRNA processing and metabolism. Drawing on recent advances in microscopy, whole genome sequencing, microarrays, mass spectrometry, fluorescent detection methodologies, and RNA interference, the authors offer readily reproducible methods for the cotranscriptional processing events that occur while the mRNA is engaged with elongating RNA polymerase II, with splicing and its biochemical analysis and with alternative splicing. Additional methods cover mRNA export, the recovery and analysis of mRNP complexes, cytoplasmic translation, mRNA degradation in vivo and in vitro, and the controversial concept of nuclear translation. A variety of model organisms are used, including yeast, Drosophila, Xenopus, mice, plants, and cultured mammalian cells. Each proven protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Timely and authoritative, mRNA Processing and Metabolism: Methods and Protocols provides a powerful collection of novel techniques that are not only effective, but also immediately applicable to current problems in many areas of biological research.
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Includes bibliographical references and index.

Using chromatin immunoprecipitation to map cotranscriptional mRNA processing in saccharomyces cerevisiae / Michael-Christopher Keogh and Stephen Buratowski -- Identifying phosphoCTD-associating proteins / Hemali P. Phatnani and Arno L. Greenleaf -- Imaging alternative splicing in living cells / Eric J. Wagner [and others] -- Tobramycin affinity tag purification of spliceosomes / Klaus Hartmuth, Hans-Peter Vornlocher and Reinhard Lührmann -- Using single-strand conformational polymorphism gel electrophoresis to analyze mutually exclusive alternative splicing / Alicia M. Celotto and Brenton R. Graveley -- Methods for the analysis of adenosine-to-inosine editing in RNA / Zuo Zhang and Gordon G. Carmichael -- Assaying nuclear messenger RNA export in human cells / Bryan R. Cullen -- RNA unwinding assay for DExD/H-Box RNA helicases / Stephanie S. Tseng-Rogenski and Tien-Hsien Chang -- Approaches for monitoring nuclear translation / Francisco J. Iborra, Dean A. Jackson, and Peter R. Cook -- Immunopurification and analysis of protein and RNA components of mRNP in mammalian cells / Fabrice Lejeune and Lynne E. Maquat.

Gene expression analysis of messenger RNP complexes / Luiz O.F. Penalva, Scott A. Tenenbaum, and Jack D. Keene -- Using the -N peptide to tether proteins to RNAs / Julie Baron-Benhamou [and others] -- An efficient system for cap- and poly(A)-dependent translation in vitro / Yuri V. Svitkin and Nahum Sonenberg -- A poly(A) tail-responsive in vitro system for cap- or IRES-driven translation from HeLa cells / Christian Thoma, Antje Ostareck-Lederer and Matthias W. Hentze -- Assessing messenger RNA decapping in cellular extracts / Naomi Bergman [and others] -- Analysis of RNA exonucleolytic activities in cellular extracts / Devi Mukherjee [and others] -- Application of ligation-mediated reverse transcription polymerase chain reaction to the identification of in vivo endonuclease-generated messenger RNA decay intermediates / Mark N. Hanson and Daniel R. Schoenberg -- Biochemical dissection of RNA silencing in plants / Guiliang Tang and Phillip D. Zamore -- RNA interference of mRNA processing factors in drosophila S2 cells / Alicia M. Celotto and Brenton R. Graveley -- RNA interference by short hairpin RNAs expressed in vertebrate cells / Gregory J. Hannon and Douglas S. Conklin.

Print version record.

MRNA processing is a key step in gene expression that effects all the proteins within the cell. In mRNA Processing and Metabolism: Methods and Protocols, world-renowned researchers bring together the latest techniques spanning the breadth of mRNA processing and metabolism. Drawing on recent advances in microscopy, whole genome sequencing, microarrays, mass spectrometry, fluorescent detection methodologies, and RNA interference, the authors offer readily reproducible methods for the cotranscriptional processing events that occur while the mRNA is engaged with elongating RNA polymerase II, with splicing and its biochemical analysis and with alternative splicing. Additional methods cover mRNA export, the recovery and analysis of mRNP complexes, cytoplasmic translation, mRNA degradation in vivo and in vitro, and the controversial concept of nuclear translation. A variety of model organisms are used, including yeast, Drosophila, Xenopus, mice, plants, and cultured mammalian cells. Each proven protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Timely and authoritative, mRNA Processing and Metabolism: Methods and Protocols provides a powerful collection of novel techniques that are not only effective, but also immediately applicable to current problems in many areas of biological research.

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