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Functional Genomics pp 307–319Cite as

Multiple-Gene Silencing Using Antisense RNAs in Escherichia coli

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 815))

Abstract

We have developed four expression vectors to express antisense RNAs (asRNAs) by which genes of interest are silenced in Escherichia coli. The vectors are all IPTG-inducible and co-transformable in any combination and target genes are silenced conditionally and concurrently. Furthermore, in order to improve silencing efficacy, the vectors are designed to express uniquely shaped antisense RNAs, named paired termini antisense RNAs (PTasRNAs). The vectors are useful for comprehensive investigation of gene function and are applicable even if the target genes are essential for cell growth. Here, we describe methods to construct PTasRNA-expressing vectors and to evaluate silencing efficacy.

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Acknowledgments

We thank the members of our research group for their assistance. This work was supported by Kato Memorial Bioscience Foundation.

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Authors and Affiliations

  1. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi,Toyohira-ku, Sapporo, 062-8517, Japan

    Nobutaka Nakashima

  2. Department of Pathology and Infectious Diseases, Royal Veterinary College, University of London, London, UK

    Shan Goh & Liam Good

  3. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan

    Tomohiro Tamura

Authors
  1. Nobutaka Nakashima
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  2. Shan Goh
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  3. Liam Good
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  4. Tomohiro Tamura
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Corresponding author

Correspondence to Nobutaka Nakashima .

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Editors and Affiliations

  1. School of Medicine, Witten/Herdecke University, Group Stockumer Str. 10 50, Witten, 58448, Germany

    Michael Kaufmann

  2. School of Medicine, Private Universität Witten/Herdecke, Witten/Herdecke University 50, Witten, 58448, Germany

    Claudia Klinger

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Nakashima, N., Goh, S., Good, L., Tamura, T. (2012). Multiple-Gene Silencing Using Antisense RNAs in Escherichia coli . In: Kaufmann, M., Klinger, C. (eds) Functional Genomics. Methods in Molecular Biology, vol 815. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-424-7_23

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  • DOI: https://doi.org/10.1007/978-1-61779-424-7_23

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-423-0

  • Online ISBN: 978-1-61779-424-7

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