title: Transcriptome-Wide Identification of Pseudouridine Modifications Using Pseudo-seq.
abstract: A diverse array of post-transcriptional modifications is found in RNA molecules from all domains of life. While the locations of RNA modifications are well characterized in abundant noncoding RNAs, modified sites in less abundant mRNAs are just beginning to be discovered. Recent work has revealed hundreds of previously unknown and dynamically regulated pseudouridines (Ψ) in mRNAs from diverse organisms. This unit describes Pseudo-seq, an efficient, high-resolution method for identification of Ψs genome-wide. This unit includes methods for isolation of RNA from S. cerevisiae, preparation of Pseudo-seq libraries from RNA samples, and identification of sites of pseudouridylation from the sequencing data. Pseudo-seq is applicable to any organism or cell type, facilitating rapid identification of novel pseudouridylation events.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Carlile
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts.
authors.forename: Maria F
authors.initials: MF
authors.lastname: Rojas-Duran
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts.
authors.forename: Wendy V
authors.initials: WV
authors.lastname: Gilbert
mesh_terms: D011560:Pseudouridine / Q000032:analysis*; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger; D011560:Pseudouridine
keywords: RNA modification; next-generation sequencing; pseudouridine