Tracking miRNA precursor metabolic products and processing sites through completely analyzing high-throughput sequencing data |
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Authors: | Li Guo Hailing Li Jiafeng Lu Qi Yang Qinyu Ge Wanjun Gu Yunfei Bai Zuhong Lu |
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Institution: | (1) State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China;(2) Key Laboratory of Child Development and Learning Science of Ministry of Education, Southeast University, Nanjing, 210096, China |
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Abstract: | The small non-coding important regulatory molecules, microRNAs (miRNAs), have been widely and deeply studied especially combining
high-throughput sequencing technologies. Here, we attempted to track detailed miRNA precursor metabolic products and gain
further insight into pre-miRNA processing by completely analyzing high-throughput sequencing data. Highly expressed miRNA
precursors could be entirely covered by various short RNAs and small RNA fragments with a hierarchical distribution. miRNAs
and some miRNA* regions were detected quite abundant short RNAs as expected, while other regions of precursors were found
shorter RNAs or small fragments with fewer sequence counts. Furthermore, we developed a method to analyze relative expression
levels of special RNA classes according to divergence of 5′ and 3′ ends, respectively. Generally, there were several quite
abundant RNA classes from a given miRNA locus, which suggested dominant cleavage sites of Drosha and Dicer during pre-miRNA
processing. Compared with 3′ end, dominant cleavage site in 5′ end always focused on a specific position, which ensured conservation
of the identity of miRNA (5′-seed sequence, nucleotides 2–8). Overall, a comprehensive analysis of sequencing data can be
used to track pre-miRNA metabolic products and mechanism of pre-miRNA processing and metabolism. |
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