Efficient and quantitative high-throughput transfer RNA sequencing

Details

• Alternative Title:
  Nat Methods
• Personal Author:
  Zheng, Guanqun ; Qin, Yidan ; Clark, Wesley C. ; Dai, Qing ; Yi, Chengqi ; He, Chuan ; Lambowitz, Alan M. ; Pan, Tao
• Description:
  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.
• Subjects:
  Algorithms Base Sequence Gene Library HEK293 Cells High-Throughput Nucleotide Sequencing Humans Molecular Sequence Data RNA, Transfer
• Source:
  Nat Methods. 12(9):835-837
• Pubmed ID:
  26214130
• Pubmed Central ID:
  PMC4624326
• Document Type:
  Journal Article
• Funding:
  T32GM007183/GM/NIGMS NIH HHSUnited States/ ; DP1 GM105386/GM/NIGMS NIH HHSUnited States/ ; K01 HG006699/HG/NHGRI NIH HHSUnited States/ ; R01 GM037949/GM/NIGMS NIH HHSUnited States/ ; R37 GM037951/GM/NIGMS NIH HHSUnited States/ ; K01HG006699/HG/NHGRI NIH HHSUnited States/ ; GM37951/GM/NIGMS NIH HHSUnited States/ ; R01 GM037951/GM/NIGMS NIH HHSUnited States/ ; R01GM37949/GM/NIGMS NIH HHSUnited States/ ; DP1GM105386/DP/NCCDPHP CDC HHSUnited States/ ; T32 GM007183/GM/NIGMS NIH HHSUnited States/
• Volume:
  12
• Issue:
  9
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:ca01903340cd97f59d93f066c35d05314288f90ed1da84e02ecdeb269fd56128
• Download URL:
  https://stacks.cdc.gov/view/cdc/36311/cdc_36311_DS1.pdf
• File Type:
  [PDF - 209.94 KB ]