Efficient and quantitative high-throughput tRNA sequencing

被引:0
|
作者
Zheng, Guanqun [1 ]
Qin, Yidan [2 ]
Clark, Wesley C. [1 ]
Dai, Qing [3 ]
Yi, Chengqi [1 ]
He, Chuan [1 ,3 ,4 ,5 ]
Lambowitz, Alan M. [2 ]
Pan, Tao [1 ]
机构
[1] Univ Chicago, Dept Biochem & Mol Biol, Chicago, IL 60637 USA
[2] Univ Texas Austin, Inst Cellular & Mol Biol, Austin, TX 78712 USA
[3] Univ Chicago, Dept Chem, Chicago, IL 60637 USA
[4] Univ Chicago, Inst Biophys Dynam, Chicago, IL 60637 USA
[5] Univ Chicago, Howard Hughes Med Inst, Chicago, IL 60637 USA
基金
美国国家卫生研究院;
关键词
ESCHERICHIA-COLI; DNA-DAMAGE; OXIDATIVE DEMETHYLATION; ALKB; REPAIR; GENES; SEQ;
D O I
10.1038/NMETH.3478
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:835 / +
页数:5
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