Preface ........................................................ v
Contributors .................................................. ix
Part I. Imaging of Noncoding RNAs and Nuclear Bodies
1 Visualization of IncRNA by Single-Molecule Fluorescence
In Situ Hybridization ........................................ 3
Margaret Dunagin, Moran N. Cabili, John Rinn, and Arjun Raj
2 Super-Resolution Imaging of Nuclear Bodies by STED
Microscopy .................................................. 21
Yasushi Okada and Shinichi Nakagawa
3 High-Resolution 3D DNA FISH Using Plasmid Probes and
Computational Correction of Optical Aberrations to Study
Chromatin Structure at the Sub-megabase Scale ............... 37
Luca Giorgetti, Tristan Piolot, and Edith Heard
4 Time-Lapse Imaging of Nuclear Bodies ........................ 55
Saskia Hutten, Samuel Swift, and Angus I. Lamond
5 Visualization of Nucleic Acids with Synthetic Exciton-
Controlled Fluorescent Oligonucleotide Probes ............... 69
Dan Ohtan Wang and Akimitsu Okamoto
6 Live CLEM Imaging to Analyze Nuclear Structures at High
Resolution .................................................. 89
Tokuko Haraguchi, Hiroko Osakada, and Takako Koujin
7 Ultrastructural Analysis of Nuclear Bodies Using Electron
Microscopy ................................................. 105
Sylvie Souquere and Gérard Pierron
8 Analyses of Nuclear Proteins and Nucleic Acid Structures
Using Atomic Force Microscopy .............................. 119
Jamie L. Gilmore, Aiko Yoshida, Hirohide Takahashi,
Katashi Deguchi, Toshiro Kobori, Emilie Louvet, Masahiro
Kumeta, Shige H. Yoshimura, and Kunio Takeyasu
9 Genome-Wide Co-Localization Screening of Nuclear Body
Components Using a Fluorescently Tagged FLI cDNA Clone
Library .................................................... 155
Tetsuro Hirose and Naoki Goshima
Part II. Biochemical Analysis of Noncoding RNAs and Nuclear
Bodies
10 Purification of Specific Chromatin Regions Using
Oligonucleotides:
Capture Hybridization Analysis of RNA Targets ( CHART) ..... 167
Christopher P. Davis and Jason A. West
11 RNA Antisense Purification (RAP) for Mapping RNA
Interactions with Chromatin ................................ 183
Jesse Engreitz, Eric S. Lander, and Mitchell Guttman
12 In Situ Dissection of RNA Functional Subunits by Domain-
Specific Chromatin Isolation by RNA Purification (dChIRP) .. 199
Jeffrey J. Quinn and Howard Y. Chang
13 Extracting, Enriching, and Identifying Nuclear Body Sub-
Complexes Using Label-Based Quantitative Mass
Spectrometry ............................................... 215
Archa Fox, Virja Mehta, Severine Boulon, and Laura
Trinkle-Mulcahy
14 Studying RNA-Binding Protein Interactions with Target
mRNAs in Eukaryotic Cells: Native Ribonucleoprotein
Immunoprecipitation (RIP) Assays ........................... 239
Joseph A. Cozzitorto, Masaya Jimbo, Saswati Chand,
Fernando Blanco, Shruti Lal, Melissa Gilbert, Jordan
M. Winter, Myriam Gorospe, and Jonathan R. Brody
15 Cross-Linking and Immunoprecipitation of Nuclear
RNA-Binding Proteins ....................................... 247
Quan Li, Yuri Ventura, and Yukio Kawahara
16 Purification of Noncoding RNA and Bound Proteins
Using FLAG Peptide-Conjugated Antisense-Oligonucleotides ... 265
Shungo Adachi and Tohru Natsume
Part III. Cell and Molecular Biological Approach Toward
the Analysis of Noncoding RNAs and Nuclear Bodies
17 MMCT-Mediated Chromosome Engineering Technique Applicable
to Functional Analysis of IncRNA and Nuclear Dynamics ...... 277
Makiko Meguro-Horike and Shin-ichi Horike
18 Reconstitution of Nucleocytoplasmic Transport Using
Digitonin-Permeabilized Cells .............................. 291
Shingo Kose, Tomoko Funakoshi, and Naoko Imamoto
19 Genome-Wide Analysis of Long Noncoding RNA Turnover ........ 305
Hidenori Tani, Naoto Imamachi, Rena Mizutani,
Katsutoshi Imamura, Yeondae Kwon, Satoru Miyazaki, Sho
Maekawa, Yutaka Suzuki, and Nobuyoshi Akimitsu
20 Knockdown of Nuclear-Retained Long Noncoding RNAs
Using Modified DNA Antisense Oligonucleotides .............. 321
Xinying Zong, Lulu Huang, Vidisha Tripathi, Raechel
Peralta, Susan M. Freier, Shuling Guo, and Kannanganattu
V. Prasanth
21 siRNA Screening of Nuclear Proteins ........................ 333
Yuko Hasegawa and Shinichi Nakagawa
Index ......................................................... 349
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