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Removing roadblocks to deep sequencing of modified RNAs

Nature Methods volume 12, pages 821–822 (2015)Cite this article

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By removing modified nucleotides that block reverse transcriptase, two methods have now made tRNAs amenable to RNA-seq.

The use of high-throughput RNA sequencing (RNA-seq) has revealed an extraordinarily diverse population of functional RNAs present in cells. However, current approaches poorly detect RNAs that are highly modified and/or extensively folded, such as tRNAs. The complementary methods of Zheng et al.1 and Cozen et al.2 have now overcome many of the technical hurdles for globally sequencing tRNAs by biochemically removing select tRNA modifications before RNA-seq library preparation. These clever strategies have revealed new insights into tRNA regulation and should be useful for identifying other modified RNAs.

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Figure 1: Removing methylated nucleotides to sequence tRNAs.

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Acknowledgements

J.E.W. is supported by US National Institutes of Health grant R00-GM104166 and the Rita Allen Foundation.

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  1. Jeremy E. Wilusz is in the Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA.,

    Jeremy E Wilusz

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  1. Jeremy E Wilusz
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Correspondence to Jeremy E Wilusz.

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Wilusz, J. Removing roadblocks to deep sequencing of modified RNAs. Nat Methods 12, 821–822 (2015). https://doi.org/10.1038/nmeth.3516

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