Determination of enrichment factors for modified RNA in MeRIP experiments

Publikationstyp:
Zeitschriftenaufsatz
Metadaten:
Autoren
  • Kaouthar Slama
  • Aurellia Galliot
  • Franziska Weichmann
  • Jasmin Hertler
  • Regina Feederle
  • Gunter Meister
  • Mark Helm
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000462417800014&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1016/j.ymeth.2018.10.020
eISSN
1095-9130
Externe Identifier
  • Clarivate Analytics Document Solution ID: HQ4YS
  • PubMed Identifier: 30394295
ISSN
1046-2023
Zeitschrift
METHODS
Schlüsselwörter
  • MeRIP
  • Antibody
  • RNA modification
  • M(6)A/N6-methyladenosine
  • TLC
Paginierung
102 - 109
Datum der Veröffentlichung
2019
Status
Published
Titel
Determination of enrichment factors for modified RNA in MeRIP experiments
Sub types
  • Article
Ausgabe der Zeitschrift
156

Datenquelle: Web of Science (Lite)

Andere Metadatenquellen:
Autoren
  • Kaouthar Slama
  • Aurellia Galliot
  • Franziska Weichmann
  • Jasmin Hertler
  • Regina Feederle
  • Gunter Meister
  • Mark Helm
DOI
10.1016/j.ymeth.2018.10.020
ISSN
1046-2023
Zeitschrift
Methods
Sprache
en
Paginierung
102 - 109
Datum der Veröffentlichung
2019
Status
Published
Herausgeber
Elsevier BV
Herausgeber URL
http://dx.doi.org/10.1016/j.ymeth.2018.10.020
Datum der Datenerfassung
2019
Titel
Determination of enrichment factors for modified RNA in MeRIP experiments
Ausgabe der Zeitschrift
156

Datenquelle: Crossref

Abstract
In the growing field of RNA modification, precipitation techniques using antibodies play an important role. However, little is known about their specificities and protocols are missing to assess their effectiveness. Here we present a method to assess enrichment factors after MeRIP-type pulldown experiments, here exemplified with a commercial antibody against N6-methyladenosine (m<sup>6</sup>A). Testing different pulldown and elution conditions, we measure enrichment factors of 4-5 using m<sup>6</sup>A-containing mRNAs against an unmodified control of identical sequence. Both types of mRNA carry <sup>32</sup>P labels at different nucleotides, allowing their relative quantification in a mixture after digestion to nucleotides, separation by TLC and quantitative phosphorimaging of the labels.
Addresses
  • Institute of Pharmacy and Biochemistry, Johannes Gutenberg-University Mainz, Germany.
Autoren
  • Kaouthar Slama
  • Aurellia Galliot
  • Franziska Weichmann
  • Jasmin Hertler
  • Regina Feederle
  • Gunter Meister
  • Mark Helm
DOI
10.1016/j.ymeth.2018.10.020
eISSN
1095-9130
Externe Identifier
  • PubMed Identifier: 30394295
Funding acknowledgements
  • Deutsche Forschungsgemeinschaft:
Open access
false
ISSN
1046-2023
Zeitschrift
Methods (San Diego, Calif.)
Schlüsselwörter
  • Cell-Free System
  • Humans
  • Escherichia coli
  • Phosphorus Radioisotopes
  • DNA-Directed RNA Polymerases
  • Immunoglobulin G
  • Viral Proteins
  • RNA, Messenger
  • Adenosine
  • Adenosine Triphosphate
  • Chromatography, Thin Layer
  • Immunoprecipitation
  • Isotope Labeling
  • Protein Structure, Secondary
  • Protein Binding
  • Methylation
  • Models, Molecular
  • Protein Interaction Domains and Motifs
Sprache
eng
Medium
Print-Electronic
Online publication date
2018
Paginierung
102 - 109
Datum der Veröffentlichung
2019
Status
Published
Datum der Datenerfassung
2018
Titel
Determination of enrichment factors for modified RNA in MeRIP experiments.
Sub types
  • Research Support, Non-U.S. Gov't
  • Journal Article
Ausgabe der Zeitschrift
156

Datenquelle: Europe PubMed Central

Abstract
In the growing field of RNA modification, precipitation techniques using antibodies play an important role. However, little is known about their specificities and protocols are missing to assess their effectiveness. Here we present a method to assess enrichment factors after MeRIP-type pulldown experiments, here exemplified with a commercial antibody against N6-methyladenosine (m6A). Testing different pulldown and elution conditions, we measure enrichment factors of 4-5 using m6A-containing mRNAs against an unmodified control of identical sequence. Both types of mRNA carry 32P labels at different nucleotides, allowing their relative quantification in a mixture after digestion to nucleotides, separation by TLC and quantitative phosphorimaging of the labels.
Date of acceptance
2018
Autoren
  • Kaouthar Slama
  • Aurellia Galliot
  • Franziska Weichmann
  • Jasmin Hertler
  • Regina Feederle
  • Gunter Meister
  • Mark Helm
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/30394295
DOI
10.1016/j.ymeth.2018.10.020
eISSN
1095-9130
Zeitschrift
Methods
Schlüsselwörter
  • Antibody
  • M(6)A/N6-methyladenosine
  • MeRIP
  • RNA modification
  • TLC
  • Adenosine
  • Adenosine Triphosphate
  • Cell-Free System
  • Chromatography, Thin Layer
  • DNA-Directed RNA Polymerases
  • Escherichia coli
  • Humans
  • Immunoglobulin G
  • Immunoprecipitation
  • Isotope Labeling
  • Methylation
  • Models, Molecular
  • Phosphorus Radioisotopes
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Structure, Secondary
  • RNA, Messenger
  • Viral Proteins
Sprache
eng
Country
United States
Paginierung
102 - 109
PII
S1046-2023(18)30304-9
Datum der Veröffentlichung
2019
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2019
Titel
Determination of enrichment factors for modified RNA in MeRIP experiments.
Sub types
  • Journal Article
  • Research Support, Non-U.S. Gov't
Ausgabe der Zeitschrift
156

Datenquelle: PubMed

Beziehungen:
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