In Vitro Reversible Translation Control Using 纬PNA Probes

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• 作者：Taylor D. Canady ; Cheryl A. Telmer ; Stanley N. Oyaghire ; Bruce A. Armitage ; Marcel P. Bruchez
• 刊名：Journal of the American Chemical Society
• 出版年：2015
• 出版时间：August 19, 2015
• 年：2015
• 卷：137
• 期：32
• 页码：10268-10275
• 全文大小：485K
• ISSN：1520-5126

文摘

On-demand regulation of gene expression in living cells is a central goal of chemical biology and antisense therapeutic development. While significant advances have allowed regulatory modulation through inserted genetic elements, on-demand control of the expression/translation state of a given native gene by complementary sequence interactions remains a technical challenge. Toward this objective, we demonstrate the reversible suppression of a luciferase gene in cell-free translation using Watson鈥揅rick base pairing between the mRNA and a complementary 纬-modified peptide nucleic acid (纬PNA) sequence with a noncomplementary toehold. Exploiting the favorable thermodynamics of 纬PNA鈭捨砅NA interactions, the antisense sequence can be removed by hybridization of a second, fully complementary 纬PNA, through a strand displacement reaction, allowing translation to proceed. Complementary RNA is also shown to displace the bound antisense 纬PNA, opening up possibilities of in vivo regulation by native gene expression.