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Single-nucleotide-resolution mapping of m6A and m6Am throughout the transcriptome | Nature Methods
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Unique mutational signatures induced by cross-linking of m6A-specific antibodies to RNA identify m6A and m6Am residues at single-nucleotide resolution, transcriptome-wide. N6-methyladenosine (m6A) is the most abundant modified base in eukaryotic mRNA and has been linked to diverse effects on mRNA fate. Current mapping approaches localize m6A residues to transcript regions 100–200 nt long but cannot identify precise m6A positions on a transcriptome-wide level. Here we developed m6A individual-nucleotide-resolution cross-linking and immunoprecipitation (miCLIP) and used it to demonstrate that antibodies to m6A can induce specific mutational signatures at m6A residues after ultraviolet light–induced antibody-RNA cross-linking and reverse transcription. We found that these antibodies similarly induced mutational signatures at N6,2′-O-dimethyladenosine (m6Am), a modification found at the first nucleotide of certain mRNAs. Using these signatures, we mapped m6A and m6Am at single-nucleotide resolution in human and mouse mRNA and identified small nucleolar RNAs (snoRNAs) as a new class of m6A-containing non-coding RNAs (ncRNAs).
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nature portfolio german research foundation permissions reprints par-clip–based pa-m6a-seq26 method privacy policy advertising nature 485 nature photo-crosslinking-assisted m6a sequencing middle table social media brain development n6-methyladenosine modification involved high-resolution n6-methyladenosine rna n6-methyladenosine modification paired-end sequencing strategy high-resolution mass spectrometry cross-linking events present single-nucleotide-resolution mapping conventional merip-seq peaks vivo protein-rna interactions interactive tools author correspondence existing merip-seq datasets pa-m6a-seq clusters photo-cross-linking single-nucleotide-resolution map springerlink instant access underlie merip-seq peaks tools permissions merip-seq–identified regions pa-m6a-seq26 uv cross-linking institutional subscriptions read par-clip data initial miclip libraries unique miclip reads miclip-called m6a residues microrna target sites cims miclip libraries existing data sets individual nucleotide resolution individual-nucleotide resolution single-nucleotide resolution single nucleotide resolution pa-m6a-seq cits miclip–called sites protein-rna interactions nucleotide-resolution studies
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description:Unique mutational signatures induced by cross-linking of m6A-specific antibodies to RNA identify m6A and m6Am residues at single-nucleotide resolution, transcriptome-wide. N6-methyladenosine (m6A) is the most abundant modified base in eukaryotic mRNA and has been linked to diverse effects on mRNA fate. Current mapping approaches localize m6A residues to transcript regions 100â200 nt long but cannot identify precise m6A positions on a transcriptome-wide level. Here we developed m6A individual-nucleotide-resolution cross-linking and immunoprecipitation (miCLIP) and used it to demonstrate that antibodies to m6A can induce specific mutational signatures at m6A residues after ultraviolet lightâinduced antibody-RNA cross-linking and reverse transcription. We found that these antibodies similarly induced mutational signatures at N6,2â²-O-dimethyladenosine (m6Am), a modification found at the first nucleotide of certain mRNAs. Using these signatures, we mapped m6A and m6Am at single-nucleotide resolution in human and mouse mRNA and identified small nucleolar RNAs (snoRNAs) as a new class of m6A-containing non-coding RNAs (ncRNAs).
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