Efficient and quantitative high-throughput tRNA sequencing.

Zheng, Guanqun; Qin, Yidan; Clark, Wesley C; Dai, Qing; Yi, Chengqi; He, Chuan; Lambowitz, Alan M; Pan, Tao · Nat Methods · 2015

basic_science · Level V

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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.

Medical subject headings