title: Efficient and quantitative high-throughput tRNA sequencing.
abstract: Despite its biological importance, tRNA has not been adequately sequenced by standard methods because of its abundant post-transcriptional modifications and stable structure, which interfere with cDNA synthesis. We achieved efficient and quantitative tRNA sequencing in HEK293T cells by using engineered demethylases to remove base methylations and a highly processive thermostable group II intron reverse transcriptase to overcome these obstacles. Our method, DM-tRNA-seq, should be applicable to investigations of tRNA in all organisms.
authors.affiliation: Department of Biochemistry & Molecular Biology, the University of Chicago, Chicago, IL 60637, USA.
authors.forename: Guanqun
authors.initials: G
authors.lastname: Zheng
authors.affiliation: Institute for Cellular and Molecular Biology, the University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Yidan
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Department of Biochemistry & Molecular Biology, the University of Chicago, Chicago, IL 60637, USA.
authors.forename: Wesley C
authors.initials: WC
authors.lastname: Clark
authors.affiliation: Department of Chemistry, the University of Chicago, Chicago, IL 60637, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Dai
authors.affiliation: Department of Biochemistry & Molecular Biology, the University of Chicago, Chicago, IL 60637, USA.
authors.forename: Chengqi
authors.initials: C
authors.lastname: Yi
authors.affiliation: Department of Biochemistry & Molecular Biology, the University of Chicago, Chicago, IL 60637, USA.|Department of Chemistry, the University of Chicago, Chicago, IL 60637, USA.|Institute of Biophysical Dynamics, the University of Chicago, Chicago, IL 60637, USA.|Howard Hughes Medical Institute, the University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Chuan
authors.initials: C
authors.lastname: He
authors.affiliation: Institute for Cellular and Molecular Biology, the University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Alan M
authors.initials: AM
authors.lastname: Lambowitz
authors.affiliation: Department of Biochemistry & Molecular Biology, the University of Chicago, Chicago, IL 60637, USA.|Institute of Biophysical Dynamics, the University of Chicago, Chicago, IL 60637, USA.
authors.forename: Tao
authors.initials: T
authors.lastname: Pan
mesh_terms: D000465:Algorithms; D001483:Base Sequence; D015723:Gene Library; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D012343:RNA, Transfer / Q000235:genetics*
chemical_list: D012343:RNA, Transfer