Functional characterization of 2'-O-methylation and pseudouridylation guide RNAs.
Pseudouridines and 2'-O-methylated nucleotides are ubiquitous constituents of stable cellular RNAs. In eukaryotes, posttranscriptional synthesis of most pseudouridines and 2'-O-methylated nucleotides is directed by sequence-specific guide RNAs (gRNAs). In recent years, an enormous number of novel putative modification gRNAs have been identified in a broad variety of organisms by using bioinformatics and large-scale cDNA-sequencing approaches. Understanding of the function of the novel modification gRNAs as well as the functional importance of modified nucleotides requires techniques that support the site-specific detection of 2'-O-methylated nucleotides and pseudouridines. Here, we describe rapid, reverse transcription-based methods to map 2'-O-methylated nucleotides and pseudouridines in any cellular RNA.
Kiss
Tamás
T
Laboratoire de Biologie Moléculaire Eucaryote du CNRS, Université Paul Sabatier, Toulouse, France.
Jády
Beáta E
BE
D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D008745:Methylation; D009838:Oligodeoxyribonucleotides / Q000302:isolation & purification; D011560:Pseudouridine / Q000032:analysis*; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics; D012265:Ribonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D012359:tRNA Methyltransferases; D058727:RNA, Small Untranslated
D017931:DNA Primers; D009838:Oligodeoxyribonucleotides; D012265:Ribonucleotides; D011560:Pseudouridine; D012359:tRNA Methyltransferases; D058727:RNA, Small Untranslated