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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 5. — A position-specific asymmetric bulge is the determinant of a 20-nt miRNA with an 18-bp duplex region. (A) miRNAs of 20-nt with an 18-bp duplex region all possess an asymmetric bulge at neighboring positions. The 20-nt miRNA strand and the complementary strand are highlighted in red and turquoise, respectively. If a 20-nt miRNA is in the 3′ arm of a pre-miRNA, the duplex is rotated 180 degrees. Asymmetric bulges located in the 3′ half of the 20-nt miRNA complementary strand are highlighted in yellow. (B) An asymmetric bulge in the 3′ half of miR156c* is required for the formation of 20-nt ath-miR156c. The length of miR156c and miR156c* derived from the original and modified precursors transiently overexpressed in N. benthamiana was analyzed with small RNA northern blot. The migration of endogenous miR156c was determined by RNA extracted from A. thaliana seedlings (ath-seedlings). Similar experimental procedures were performed to determine miRNA length in (C) and (D). (C) An asymmetric bulge in the 3′ half of miR158a* is required for the formation of 20-nt ath-miR158a. (D) The effect of an asymmetric bulge on ath-miR156c length is position-dependent. Asymmetric bulges introduced in modified constructs are highlighted in yellow. Open arrowheads indicate the 24-nt siRNA resulted from transient overexpression of miRNA in N. benthamiana. A representative small RNA RNA gel blot derived from 3 independent transient expression assays is shown.