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Combinatorial, selective and reversible control of gene expression using oligodeoxynucleotides in a cell‐free protein synthesis system
Author(s) -
Keum JungWon,
Ahn JinHo,
Kang Taek Jin,
Kim DongMyung
Publication year - 2008
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22081
Subject(s) - gene expression , gene , messenger rna , biology , sense (electronics) , microbiology and biotechnology , protein biosynthesis , regulation of gene expression , antisense rna , chemistry , biochemistry
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.