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Highly efficient gene silencing using perfect complementary artificial miRNA targeting AP1 or heteromeric artificial miRNA targeting AP1 and CAL genes

  • Genetics and Genomics
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Abstract

Gene silencing is a useful technique for elucidating biological function of genes by knocking down their expression. Recently developed artificial microRNAs (amiRNAs) exploit an endogenous gene silencing mechanism that processes natural miRNA precursors to small silencing RNAs that target transcripts for degradation. Based on natural miRNA structures, amiRNAs are commonly designed such that they have a few mismatching nucleotides with respect to their target sites as well as within mature amiRNA duplexes. In this study, we performed an analysis in which the conventional and modified form of an amiRNA was compared side by side. We showed that the amiRNA containing 5′ mismatch with its amiRNA* and perfect complementarity to its target gene acted as a highly potent gene silencing agent against AP1, achieving a desired null mutation effect. In addition, a simultaneous silencing of two independent genes, AP1 and CAL1 was tested by employing a multimeric form of amiRNAs. Advantages and potential disadvantages of using amiRNAs with perfect complementarity to the target gene are discussed. The results presented here should be helpful in designing more specific and effective gene silencing agents.

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Acknowledgments

We owe thanks to M. Aukerman and B.-C. Yoo for helpful discussions and comments on the manuscript and H. Frick and R. Sager for careful proofreading. We also thank D. Weigel for providing pRS300 plasmid vector. This research was supported by the National Institutes of Health COBRE (P20 RR15588) and DuPont Young Professorship awarded to J.-Y. L.

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  1. Wonkeun Park

    Present address: USDA-ARS, Florence, SC, USA

Authors and Affiliations

  1. Department of Plant and Soil Sciences, Delaware Biotechnology Institute, University of Delaware, Newark, DE, 19711, USA

    Wonkeun Park, Jixian Zhai & Jung-Youn Lee

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  1. Wonkeun Park
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  2. Jixian Zhai
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  3. Jung-Youn Lee
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Correspondence to Jung-Youn Lee.

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Communicated by D. Somers.

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Park, W., Zhai, J. & Lee, JY. Highly efficient gene silencing using perfect complementary artificial miRNA targeting AP1 or heteromeric artificial miRNA targeting AP1 and CAL genes. Plant Cell Rep 28, 469–480 (2009). https://doi.org/10.1007/s00299-008-0651-5

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  • DOI: https://doi.org/10.1007/s00299-008-0651-5

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