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. 2015 Sep 18;12(9):1054–1066. doi: 10.1080/15476286.2015.1079682

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© 2015 The Author(s). Published with license by Taylor & Francis Group, LLC

© Wen-Chi Lee, Shin-Hua Lu, Ming-Hsuan Lu, Chen-Jui Yang, Shu-Hsing Wu, and Ho-Ming Chen

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 6. — The position of an asymmetric bulge relative to the putative 3′ binding pocket of DCL1 is crucial for the biogenesis of 20-nt miRNA. (A) Foldbacks of ath-MIR390a, ath-miR390a@156c, ath-MIR390a-12B, ath-MIR390a-Inv and ath-MIR390a-Inv-12B. The sequences of miR390a and miR390a* in each foldback are highlighted in red and turquoise, respectively. Red arrows indicate the directions of DCL1 processing and the nucleotides presumably associated with the putative 3′ binding pocket of DCL1 are indicated with rectangles. Asymmetric bulges which were introduced to ath-MIR390a are highlighted in yellow. (B) The precursor of ath-miR156c is sufficient for the production of artificial 20-nt ath-miR390a. (C) The introduction of an asymmetric bulge to the duplex region of ath-MIR390a failed to result in predominant expression of 20-nt miRNA. (D) An asymmetric bulge needs to be in the arm away from the putative 3′ binding pocket of DCL1 to convert ath-MIR390a into a 20-nt miRNA precursor. Amounts of total RNA loaded into each lane are indicated. Open arrowheads indicate the 24-nt siRNA resulted from transient overexpression of miRNA in N. benthamiana. A representative small RNA northern blot derived from 3 independent transient expression assays is shown.