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The Menu of Features that Define Primary MicroRNAs and Enable De Novo Design of MicroRNA Genes

Author(s)
Fang, Wenwen; Bartel, David
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Abstract
MicroRNAs (miRNAs) are small regulatory RNAs processed from stem-loop regions of primary transcripts (pri-miRNAs), with the choice of stem loops for initial processing largely determining what becomes a miRNA. To identify sequence and structural features influencing this choice, we determined cleavage efficiencies of > 50,000 variants of three human pri-miRNAs, focusing on the regions intractable to previous high-throughput analyses. Our analyses revealed a mismatched motif in the basal stem region, a preference for maintaining or improving base pairing throughout the remainder of the stem, and a narrow stem-length preference of 35 ± 1 base pairs. Incorporating these features with previously identified features, including three primary-sequence motifs, yielded a unifying model defining mammalian pri-miRNAs in which motifs help orient processing and increase efficiency, with the presence of more motifs compensating for structural defects. This model enables generation of artificial pri-miRNAs, designed de novo, without reference to any natural sequence yet processed more efficiently than natural pri-miRNAs.
Date issued
2015-09
URI
http://hdl.handle.net/1721.1/116324
Department
Massachusetts Institute of Technology. Department of Biology
Journal
Molecular Cell
Publisher
Elsevier BV
Citation
Fang, Wenwen and David P. Bartel. “The Menu of Features That Define Primary MicroRNAs and Enable De Novo Design of MicroRNA Genes.” Molecular Cell 60, 1 (October 2015): 131–145 © 2015 Elsevier Inc
Version: Author's final manuscript
ISSN
1097-2765
1097-4164

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