MicroRNA methods

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Sprache:English
Veröffentlicht: Amsterdam [u.a.] Elsevier, Acad. Press 2007
Schriftenreihe:Methods in enzymology 427
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MARC

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Datensatz im Suchindex

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adam_text Contents Contributors xi Volumes in Series xv Section I. Identifying M ¡ero R N As and Their Targets ι 1. Identification of Viral MicroRNAs 3 Christopher S. Sullivan and Adam Grundhoff 1. Introduction 3 2. Computational Prediction of Viral miRNA Candidates 7 3. Array Confirmation of Viral miRNA Candidates 11 4. Concluding Remarks 21 References 22 2. Robust Machine Learning Algorithms Predict MicroRNA Genes and Targets 25 Pål Saätrom and Ola Snove, Jr. 1. Introduction 26 2. Appropriate Use of Machine Learning 27 3. One Deterministic and One Stochastic Algorithm 33 4. miRNA Gene Prediction with Support Vector Machines 37 5. miRNA Target Prediction with Boosted Genetic Programming 43 6. Summary 46 Acknowledgments 46 References 46 3. Identification of Virally Encoded MicroRNAs 51 Sébastien Pfeffer 1. Introduction 52 2. Purification of Rnb (1-249) ar|d Adenylation of 3 Adapter Oligonucleotide 55 3. Isolation of the Small RNA Fraction 55 4. Ligation of the Purified Small RNA to Adenylated 3 Adapter 56 5. Ligation of the Small RNA-3 Adapter to the 5 Adapter 56 6. Reverse Transcription of the Final Ligation Product 57 vi Contents 7. First PCR Amplification of the cDNA 57 8. Pme / Digestion of the PCR Product 58 9. Second PCR Amplification 58 10. Ban / Digestion of the Second PCR Product 59 11. Concatamerization of the Ban/-Digested DNA 59 12. End Tailing of Concatamers and Cloning into T/A Vector 59 13. Sequencing and Annotation of the Library 60 14. Concluding Remarks 60 References 61 4. Computational Methods for MicroRNA Target Prediction 65 Yuka Watanabe, Masaru Tornita, and Akio Kanai 1. Introduction 66 2. Principles of miRNA Target Recognition 69 3. Resources for Analysis of miRNA Target Genes 73 4. Software Useful for miRNA Target Prediction 74 5. Original Strategies for Prediction of miRNA Target Genes 79 6. Validation of Computational Predictions 81 7. Concluding Remarks 82 Acknowledgments 83 References 83 Section II. MicroRNA Expression, Maturation, and Functional Analysis 87 5. In Vitro and In Vivo Assays for the Activity of Drosha Complex 89 Yoontae Lee and V. Narry Kim 1. Introduction 89 2, Assay Methods 94 Acknowledgments 105 References 105 6. Microarray Analysis of miRNA Gene Expression 107 J. Michael Thomson, Joel S. Parker, and Scott M. Hammond 1. Introduction 107 2. Overview of miRNA Biogenesis and Effector Pathways 109 3. miRNA Expression Analysis Strategies 110 4. Considerations for miRNA Microarrays 111 5. Data Analysis and Interpretation 112 Contents vii 6. Validation Strategies 113 7. rníRNA Microarray Protocol 113 8. Data Analysis 118 Acknowledgments 119 References 119 7. Cloning and Detecting Signature MicroRNAs from Mammalian Cells 123 Guihua Sun, Haitang Li, and John J. Rossi 1. Introduction 124 2. miRNA Cloning 126 3. miRNA Identification 133 4. Northern Hybridization to Verify In Vivo Expression of miRNA 134 Acknowledgments 136 References 136 8. Approaches for Studying MicroRNA and Small Interfering RNA Methylation In Vitro and In Vivo 139 Zhiyong Yang, Giedrius Vilkaitis, Bin Yu, Saulius Klimašauskas, and Xuemei Chen 1. Introduction 140 2. Expression and Purification of Recombinant HENi Proteins 141 3. Small RNA Methyttransferase Assays with Recombinant HENi Proteins 144 4. Reverse-Phase HPLC Analysis to Determine the Position of the Methyl Group in Products of HENi-Catalyzed Reactions 148 5. Immunoprecipitation and HENi Activity Assay 149 6. Analysis of the In Vivo Methylation Status of miRNAs and siRNAs 151 7. Concluding Remarks 152 Acknowledgments 152 References 153 9. Analysis of Small RNA Profiles During Development 155 Toshiaki Watanabe, Yasushi Totoki, Hiroyuki Sasaki, Naojiro Minami, and Hiroshi Imai 1. Introduction 156 2. Preparation of Low-Molecular-Weight RNA and Urea-Polyacrylamide Gel Electrophoresis (Urea-PAGE) 158 3. Cloning of Small RNAs 161 viii Contents 4. Classification of Small RNAs 163 5. Northern Blot Analysis 165 References 167 10. Dissecting MicroRNA-mediated Gene Regulation and Function in Т -Cell Development 171 Tin Ky Mao and Chang-Zheng Chen 1. Introduction 172 2. Characterizing miRNA Expression During Т -Cell Development 172 3. Retroviral Constructs for miRNA Expression 178 4. Investigating miRNA Function in Т -Cell Development 180 5. Identification and Validation of Functionally Relevant miRNA Target Genes 183 6. Materials and Reagents 187 Acknowledgments 188 References 188 Section III. MicroRNAs and Disease 191 11. Investigation of MicroRNAs Alterations in Leukémiás and Lymphomas 193 George Adrian Calin and Carlo Maria Croce 1. MicroRNA Alterations are Involved in the Initiation and Progression of Every Type of Human Cancer 194 2. Genome-Wide MicroRNA Profiling by Microarray 195 3. Identification and Validation of Targets for Differentially Expressed miRNAs 205 Acknowledgments 210 References 210 12. Discovery of Pathogen-Regulated Small RNAs in Plants 215 Surekha Katiyar-Agarwal and Hailing Jin ι. Introduction 216 2. Sequencing-Based Approaches for the Discovery of Pathogen-Regulated Small RNAs 217 3. Hybridization-Based Approaches of Identifying and Validating Pathogen-lnducible Small RNAs 219 4. Concluding Remarks 225 References 226 Contents ix 13. Protocols for Expression and Functional Analysis of Viral MicroRNAs 229 Eva Gottwein and Bryan R. Cullen 1. Introduction 230 2. miRNA Expression Cassettes 232 3. Vector Systems for Stable Delivery 233 4. Protocol: Generation of miRNA-Expressing Cell Lines Using pNL-SIN-CMV-BLR-Based miRNA Expression Vectors 235 5. Indicator Assays Establish miRNA Activity 237 6. Protocol: Preparation of Virus Mixes and Indicator Assay 237 7. Concluding Remarks 240 Acknowledgments 240 References 240 Author Index 24s Subject Index 257
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physical XL, 261 S., [4] Bl. Ill., graph. Darst.
publishDate 2007
publishDateSearch 2007
publishDateSort 2007
publisher Elsevier, Acad. Press
record_format marc
series Methods in enzymology
series2 Methods in enzymology
spellingShingle MicroRNA methods
Methods in enzymology
MicroRNAs
RNA, Messenger physiology
Sequence Analysis, RNA
Small interfering RNA
Labortechnik (DE-588)4123602-6 gnd
Small RNA (DE-588)7501213-3 gnd
subject_GND (DE-588)4123602-6
(DE-588)7501213-3
(DE-588)4143413-4
title MicroRNA methods
title_auth MicroRNA methods
title_exact_search MicroRNA methods
title_full MicroRNA methods ed. by John J. Rossi ...
title_fullStr MicroRNA methods ed. by John J. Rossi ...
title_full_unstemmed MicroRNA methods ed. by John J. Rossi ...
title_short MicroRNA methods
title_sort microrna methods
topic MicroRNAs
RNA, Messenger physiology
Sequence Analysis, RNA
Small interfering RNA
Labortechnik (DE-588)4123602-6 gnd
Small RNA (DE-588)7501213-3 gnd
topic_facet MicroRNAs
RNA, Messenger physiology
Sequence Analysis, RNA
Small interfering RNA
Labortechnik
Small RNA
Aufsatzsammlung
url http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=015957682&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
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