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Inhibiting replication of begomoviruses using artificial zinc finger nucleases that target viral-conserved nucleotide motif

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Abstract

Geminiviridae consists of a large group of single-stranded DNA viruses that cause tremendous losses worldwide. Frequent mixed infection and high rates of recombination and mutation allow them to adapt rapidly to new hosts and overcome hosts’ resistances. Therefore, an effective strategy able to confer plants with resistance against multiple begomoviruses is needed. In the present study, artificial zinc finger proteins were designed based on a conserved sequence motif of begomoviruses. DNA-binding affinities and specificities of these artificial zinc fingers were evaluated using electrophoretic mobility shift assay. Artificial zinc finger nuclease (AZFNs) were then constructed based on the ones with the highest DNA-binding affinities. In vitro digest and transient expression assay showed that these AZFNs can efficiently cleave the target sequence and inhibit the replication of different begomoviruses. These results suggest that artificial zinc finger protein technology may be used to achieve resistance against multiple begomoviruses.

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Acknowledgement:

This study was supported by the National Natural Science Foundation of China (Project Nos. 31101417 and 31101415), the Natural Science Foundation of Zhejiang Province (Project Nos: Y3110175 and Y3110277) and the Scientific Research Foundation of Hangzhou Normal University (Project No: 2351001020).

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Authors and Affiliations

  1. College of Life and Environmental Science, Hangzhou Normal University, Hangzhou, 310036, Zhejiang, People’s Republic of China

    Wei Chen, Yongsheng Qian, Yuqiang Sun & Xiaofei Cheng

  2. College of Agricultural and Food Science, Zhejiang Agricultural and Forestry University, Linan, 311300, Zhejiang, People’s Republic of China

    Xiaoyun Wu

  3. Key Laboratory of Soybean Biology of Ministry of Education, College of Agriculture, Northeast Agricultural University, Harbin, 150030, Heilongjiang, People’s Republic of China

    Xiaoxia Wu

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  1. Wei Chen
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  2. Yongsheng Qian
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  4. Yuqiang Sun
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  5. Xiaoxia Wu
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  6. Xiaofei Cheng
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Correspondence to Xiaofei Cheng.

Additional information

Wei Chen and Yongsheng Qian contributed equally to this study.

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Chen, W., Qian, Y., Wu, X. et al. Inhibiting replication of begomoviruses using artificial zinc finger nucleases that target viral-conserved nucleotide motif. Virus Genes 48, 494–501 (2014). https://doi.org/10.1007/s11262-014-1041-4

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  • DOI: https://doi.org/10.1007/s11262-014-1041-4

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