title: Genome-Wide Location Analyses of N6-Methyladenosine Modifications (m6A-Seq).
abstract: N6-methyladenosine-sequencing (m6A-seq) is a critical tool to obtain an unbiased genome-wide picture of m6A sites of modification at high resolution. It allows the study of the impact of various perturbations on m6A modification distribution and the study of m6A functions. Herein, we describe the m6A-seq protocol, which entails RNA immunoprecipitation (RIP) performed on fragmented poly(A) RNA utilizing anti-m6A antibodies. The captured/enriched m6A positive RNA fragments are subsequently sequenced by RNA-seq in parallel with background control non-immunoprecipitated input RNA fragments. Analyses reveal peaks of m6A enrichment containing sites of modifications analogous to chromatin modification immunoprecipitation experiments.
authors.affiliation: Gastrointestinal Unit, 55 Fruit Street, Jackson 828, Boston, MA, 02114, USA. molinie.benoit@gmail.com.
authors.forename: Benoit
authors.initials: B
authors.lastname: Molinie
authors.affiliation: Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, 55 Fruit Street, Jackson 828, Boston, MA, 02114, USA. cgiallourakis@mgh.harvard.edu.
authors.forename: Cosmas C
authors.initials: CC
authors.lastname: Giallourakis
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives*; D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling; D016678:Genome; D008745:Methylation; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017423:Sequence Analysis, RNA; D059467:Transcriptome; D057188:Workflow
chemical_list: D012333:RNA, Messenger; D012313:RNA; C010223:N-methyladenosine; D000241:Adenosine
keywords: Epitranscriptome; Genome-wide; METTL14; METTL3; N6-Methyladenosine; m6A-Seq