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Nucleic Acids Research Advance Access originally published online on June 26, 2008
Nucleic Acids Research 2008 36(13):4277-4285; doi:10.1093/nar/gkn388
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Nucleic Acids Research, 2008, Vol. 36, No. 13 4277-4285
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

Dispatched Homolog 2 is targeted by miR-214 through a combination of three weak microRNA recognition sites

Nan Li1, Alex S. Flynt1, H. Rosemary Kim2, Lilianna Solnica-Krezel1 and James G. Patton1,*

1Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA and 2MRC Centre for Development and Biomedical Genetics, University of Sheffield, Sheffield, UK

*To whom correspondence should be addressed. Tel: 615-322-4738; Fax: 615-343-6707; Email: james.g.patton{at}vanderbilt.edu

Received May 7, 2008. Revised May 30, 2008. Accepted June 2, 2008.

MicroRNAs (miRNAs) regulate gene expression by inhibiting translation of target mRNAs through pairing with miRNA recognition elements (MREs), usually in 3' UTRs. Because pairing is imperfect, identification of bona fide mRNA targets presents a challenge. Most target recognition algorithms strongly emphasize pairing between nucleotides 2–8 of the miRNA (the ‘seed’ sequence) and the mRNA but adjacent sequences and the local context of the 3' UTR also affect targeting. Here, we show that dispatched 2 is a target of miR-214. In zebrafish, dispatched 2 is expressed in the telencephalon and ventral hindbrain and is essential for normal zebrafish development. Regulation of dispatched 2 by miR-214 is via pairing with three, noncanonical, weak MREs. By comparing the repression capacity of GFP reporters containing different dispatched 2 sequences, we found that a combination of weak sites, which lack canonical seed pairing, can effectively target an mRNA for silencing. This finding underscores the challenge that prediction algorithms face and emphasizes the need to experimentally validate predicted MREs.


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[Abstract] [Full Text] [PDF]


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