Get access

Combinatorial, selective and reversible control of gene expression using oligodeoxynucleotides in a cell-free protein synthesis system

Authors

  • Jung-Won Keum,

    1. School of Chemical and Biological Engineering, College of Engineering, Seoul National University, Seoul, Korea
    Current affiliation:
    1. Department of Chemical Engineering, University of Massachusetts, Amherst, MA 01003.
    Search for more papers by this author
  • Jin-Ho Ahn,

    1. Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea; telephone: +82-42-821-5899; fax: +82-42-823-7692
    Search for more papers by this author
  • Taek Jin Kang,

    1. School of Chemical and Biological Engineering, College of Engineering, Seoul National University, Seoul, Korea
    Current affiliation:
    1. Research Center for Advanced Science and Technology, The University of Tokyo, 153-8904 Tokyo, Japan.
    Search for more papers by this author
  • Dong-Myung Kim

    Corresponding author
    1. Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea; telephone: +82-42-821-5899; fax: +82-42-823-7692
    • Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea; telephone: +82-42-821-5899; fax: +82-42-823-7692.
    Search for more papers by this author

  • Jung-Won Keum and Jin-Ho Ahn contributed equally to this work.

Abstract

Herein we describe the methods for selective and reversible regulation of gene expression using antisense oligodeoxynucleotides (ODNs) in a cell-free protein synthesis system programmed with multiple DNAs. Either a complete shut down or controlled level of gene expression was attained through the antisense ODN-mediated regulation of mRNA stability in the reaction mixture. In addition to the primary control of gene expression, we also demonstrate that the inhibition of protein synthesis can be reversed by using an anti-antisense ODN sequence that strips the antisense ODN off the target sequence of mRNA. As a result, sequential additions of the antisense and anti-antisense ODNs enabled the stop-and-go expression of protein molecules. Through the on-demand regulation of gene expression, presented results will provide a versatile platform for the analysis and understanding of the complicated networks of biological components. Biotechnol. Bioeng. 2009;102: 577–582. © 2008 Wiley Periodicals, Inc.

Get access to the full text of this article

Ancillary