PAR-CLIP (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation): a step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins.

Spitzer, Jessica; Hafner, Markus; Landthaler, Markus; Ascano, Manuel; Farazi, Thalia; Wardle, Greg; Nusbaum, Jeff; Khorshid, Mohsen; Burger, Lukas; Zavolan, Mihaela; Tuschl, Thomas

Spitzer, Jessica; Hafner, Markus; Landthaler, Markus; Ascano, Manuel; Farazi, Thalia; Wardle, Greg; Nusbaum, Jeff; Khorshid, Mohsen; Burger, Lukas; Zavolan, Mihaela; Tuschl, Thomas.

Afiliação

Spitzer J; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA; These authors contributed equally to this work.
Hafner M; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA; These authors contributed equally to this work.
Landthaler M; Present address: Berlin Institute for Medical Systems Biology, Max-Delbruck-Center for Molecular Medicine, Berlin, Germany.
Ascano M; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA.
Farazi T; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA.
Wardle G; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA.
Nusbaum J; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA.
Khorshid M; Biozentrum der Universität Basel and Swiss Institute of Bioinformatics (SIB), Basel, Switzerland.
Burger L; Biozentrum der Universität Basel and Swiss Institute of Bioinformatics (SIB), Basel, Switzerland.
Zavolan M; Biozentrum der Universität Basel and Swiss Institute of Bioinformatics (SIB), Basel, Switzerland.
Tuschl T; Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY, USA. Electronic address: ttuschl@rockefeller.edu.

Methods Enzymol ; 539: 113-61, 2014.

Article em En | MEDLINE | ID: mdl-24581442

Assuntos

Proteínas de Ligação a RNA/metabolismo; RNA/metabolismo; Animais; Sítios de Ligação; Células Cultivadas; Endopeptidase K/química; Humanos; Imunoprecipitação; Processos Fotoquímicos; Proteólise; RNA/química; RNA/isolamento & purificação; Proteínas de Ligação a RNA/química; Proteínas de Ligação a RNA/isolamento & purificação; Transcriptoma; Raios Ultravioleta

Palavras-chave

3'- and 5'-adapter ligation; Cell lysate preparation; Crosslinking and immunoprecipitation (CLIP) protocol; Immunoprecipitation and second RNase T1 digestion; PCR amplification of cDNA library; Posttranscriptional regulation (PTR); Protein G magnetic beads; Proteinase K digestion; UV crosslinking of 4-thiouridine-labeled cells; cDNA library preparation; cDNA library preparation/reverse transcription

Texto completo

Imprimir

XML

PubMed Links

Buscar no Google

Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA / Proteínas de Ligação a RNA Idioma: En Ano de publicação: 2014 Tipo de documento: Article

Texto completo

Imprimir

XML

PubMed Links

Buscar no Google

Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA / Proteínas de Ligação a RNA Idioma: En Ano de publicação: 2014 Tipo de documento: Article