AI Article Synopsis
- Researchers explored the role of 5-hydroxymethylcytidine (hm5C) in RNA, noting its importance in cell development but lacking detection methods to study it further.
- They introduced a new sequencing technique that converts hm5C into trihydroxylated thymine (thT), which helps detect its presence in RNA.
- The study successfully combined this chemical conversion with standard sequencing methods to analyze hm5C sites in RNA, enhancing our understanding of its biological functions.
Article Abstract
While 5-hydroxymethylcytidine in RNA (hm5C) is associated with cellular development and differentiation, its distribution and biological function remain largely unexplored because suitable detection methods are lacking. Here, we report a base-resolution sequencing method for hm5C in RNA by applying peroxotungstate-mediated chemical conversion of hm5C to trihydroxylated thymine (thT). Reverse transcription by SuperScript III terminated at the thT site, probably because of its unnatural nucleobase structure producing truncated cDNA. Consequently, base-resolution analysis of the hm5C sites in RNA was achieved with both Sanger sequencing and Illumina sequencing analysis by comparing sequencing data before and after peroxotungstate treatment.
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| http://dx.doi.org/10.1039/d1ob00995h | DOI Listing |
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