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Comparative Studies of Various Artificial microRNA Expression Vectors for RNAi in Mammalian Cells

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Abstract

Artificial microRNA (amiRNA) has recently become an important RNA interference (RNAi) technology for gene therapy and gene function studies. Here nine expression strategies were employed to construct plasmid vectors expressing amiRNA (amiR-Fluc) against firefly luciferase (Fluc). Our results indicate that all nine vectors can successfully produce mature amiR-Fluc and specifically suppress the expression of Fluc, although the RNAi efficiency in different mammalian cells displays obvious differences. Among these nine vectors, three can efficiently co-express DsRed reporter gene linked with amiR-Fluc cassette. Moreover, the recommended number of concatenated amiRNAs in a multi-amiRNA expression vector should not be more than four, and the relative position of an amiRNA in the multi-amiRNA expression vector has no apparent influence on its RNAi activity. In summary, all these results described here provide valuable information for the rational design and application of amiRNA expression vector.

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Acknowledgments

This study was supported by China “973” Program (#2006CB504305 and #2010CB911800), the National Natural Science Foundation of China (#30921001 and #30925003), and National Special Research Program of Major Infectious Diseases (#2008ZX10001–002).

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Authors and Affiliations

  1. State Key Laboratory of Virology and The Modern Virology Research Centre, College of Life Sciences, Wuhan University, Wuhan, 430072, People’s Republic of China

    Tao Hu, Qiong Fu, Ye Liu, Musarat Ishaq, Junwei Li, Li Ma & Deyin Guo

  2. Laboratory of Animal Center, Zhengzhou University, Zhengzhou, 450001, People’s Republic of China

    Tao Hu

  3. Department of Pathophysiology, Basic Medical College, Zhengzhou University, Zhengzhou, 450001, People’s Republic of China

    Ping Chen

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  1. Tao Hu
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  2. Ping Chen
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  3. Qiong Fu
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  4. Ye Liu
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  5. Musarat Ishaq
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  7. Li Ma
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Correspondence to Deyin Guo.

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Hu, T., Chen, P., Fu, Q. et al. Comparative Studies of Various Artificial microRNA Expression Vectors for RNAi in Mammalian Cells. Mol Biotechnol 46, 34–40 (2010). https://doi.org/10.1007/s12033-010-9264-7

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