Pseudo-Seq: Genome-Wide Detection of Pseudouridine Modifications in RNA.

摘要: RNA molecules contain a variety of chemically diverse, posttranscriptionally modified bases. The most abundant base found in cellular RNAs, pseudouridine (Ψ), has recently been mapped to hundreds sites mRNAs, many which are dynamically regulated. Though the landscape determined only few cell types and growth conditions, enzymes responsible for mRNA pseudouridylation universally conserved, suggesting novel pseudouridylated remain be discovered. Here, we present Pseudo-seq, technique that allows identification genome-wide with single-nucleotide resolution. In this chapter, provide detailed description Pseudo-seq. We include protocols isolation from Saccharomyces cerevisiae, Pseudo-seq library preparation, data analysis, including descriptions processing mapping sequencing reads, computational pseudouridylation, assignment specific synthases. approach presented here is readily adaptable any or tissue type high-quality can isolated. Identification an important first step elucidating regulation functions these modifications.