Efficient and quantitative high-throughput tRNA sequencing

Nat Methods. 2015 Sep;12(9):835-837. doi: 10.1038/nmeth.3478. Epub 2015 Jul 27.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms*
  • Base Sequence
  • Gene Library*
  • HEK293 Cells
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Molecular Sequence Data
  • RNA, Transfer / genetics*


  • RNA, Transfer