Abstract
Recent technological advances have allowed development of increasingly complex systems for in vitro evolution. Here, we describe an in vitro autogene composed of a self-amplifying T7 RNA polymerase system. Functional autogene templates in cell-free lysate produce T7 RNA polymerase, which amplifies the autogene genetic information through a positive feedback architecture. Compartmentalization of individual templates within a water-in-oil emulsion links genotype and phenotype, allowing evolution.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Bacteriophage T7 / enzymology
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Bacteriophage T7 / genetics
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DNA-Directed RNA Polymerases / chemistry
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DNA-Directed RNA Polymerases / genetics*
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Directed Molecular Evolution*
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Feedback, Physiological
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Genes, Viral
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Nucleic Acid Conformation
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RNA, Messenger / chemistry
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RNA, Messenger / genetics
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RNA, Viral / chemistry
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RNA, Viral / genetics
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Synthetic Biology
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Viral Proteins / chemistry
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Viral Proteins / genetics*
Substances
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RNA, Messenger
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RNA, Viral
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Viral Proteins
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RNA polymerase SP6
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bacteriophage T7 RNA polymerase
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DNA-Directed RNA Polymerases