Identification of a selective DNA ligase for accurate recognition and ultrasensitive quantification of N6 -methyladenosine in RNA at one-nucleotide resolution

Chemical Science
Weiliang LiuZhengping Li

Abstract

N6-Methyladenosine (m6A) is the most frequent post-transcriptional modification in RNA, and it plays a critical role in biological processes. The functions of m6A remain largely unexplored due to a lack of highly sensitive methods to quantitatively determine the m6A modification fraction at a precise location. Here, we first reveal that T3 DNA ligase has significant selectivity towards the m6A modification. On the basis of the new finding, we establish an ultrasensitive quantitation assay for accurately determining m6A at one-nucleotide resolution in RNA. With the proposed assay, as low as 4 fM RNA containing m6A can be determined and the selectivity is up to 54.1-fold to discriminate m6A against unmodified adenosine (A). The sensitivity has been improved about 106-fold so the proposed method can be successfully employed to accurately determine m6A in real biological samples, even in low abundance RNA.

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Citations

Oct 23, 2018·Chemistry : a European Journal·Katja Hartstock, Andrea Rentmeister
Sep 15, 2019·Nature Reviews. Molecular Cell Biology·Sara ZaccaraSamie R Jaffrey
Apr 10, 2020·The Journal of Biological Chemistry·Magda BujnowskaJingyi Fei
May 7, 2020·Chemistry & Biodiversity·Shixi YangXiang Zhou
Dec 12, 2019·Nature Communications·Casslynn W Q KohW S Sho Goh
Apr 29, 2020·Nature Chemical Biology·Xiao ShuJianzhao Liu
Sep 7, 2018·Open Biology·Anna Ovcharenko, Andrea Rentmeister
Jul 10, 2019·Science Advances·Zhang ZhangGuan-Zheng Luo
Jul 3, 2019·RNA·Kristina RauAndrea Rentmeister
Dec 8, 2020·Essays in Biochemistry·Ye Wang, Guifang Jia
Mar 9, 2021·Current Opinion in Chemical Biology·Jie CaoJianzhao Liu
Jun 27, 2021·Methods : a Companion to Methods in Enzymology·Hong-Xuan ChenGuan-Zheng Luo
Jul 3, 2021·International Journal of Molecular Sciences·Vilbert SikorskiEsko Kankuri
Nov 13, 2020·Biotechnology Advances·Hong-Xiang ZhengNa Sui
Oct 17, 2020·Journal of Molecular Cell Biology·Xiao-Min LiuJun Zhou
Sep 14, 2018·Analytical Chemistry·Bei ChenYu-Qi Feng

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Methods Mentioned

BETA
immunoprecipitation
MeRIP-seq
electrophoresis
PCR
nucleic acid amplification

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