Jung-Won Keum and Jin-Ho Ahn contributed equally to this work.
Combinatorial, selective and reversible control of gene expression using oligodeoxynucleotides in a cell-free protein synthesis system†
Article first published online: 8 AUG 2008
Copyright © 2008 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 102, Issue 2, pages 577–582, 1 February 2009
How to Cite
Keum, J.-W., Ahn, J.-H., Kang, T. J. and Kim, D.-M. (2009), Combinatorial, selective and reversible control of gene expression using oligodeoxynucleotides in a cell-free protein synthesis system. Biotechnol. Bioeng., 102: 577–582. doi: 10.1002/bit.22081
- Issue published online: 17 DEC 2008
- Article first published online: 8 AUG 2008
- Accepted manuscript online: 8 AUG 2008 12:00AM EST
- Manuscript Accepted: 30 JUL 2008
- Manuscript Revised: 4 JUL 2008
- Manuscript Received: 21 MAR 2008
- Korea Science & Engineering Foundation. Grant Number: R01-2007-000-20980-0
- cell-free protein synthesis;
- synthetic biology;
- antisense oligodeoxynucleotide;
- anti-antisense oligodeoxynucleotide
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.