An in vitro autogene

Eric A Davidson; Adam J Meyer; Jared W Ellefson; Matthew Levy; Andrew D Ellington

doi:10.1021/sb3000113

An in vitro autogene

ACS Synth Biol. 2012 May 18;1(5):190-6. doi: 10.1021/sb3000113. Epub 2012 May 2.

Authors

Eric A Davidson¹, Adam J Meyer, Jared W Ellefson, Matthew Levy, Andrew D Ellington

Affiliation

¹ Department of Chemistry and Biochemistry, Institute for Cell and Molecular Biology, Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX 78712, USA.

PMID: 23651157
DOI: 10.1021/sb3000113

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

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Bacteriophage T7 / enzymology
Bacteriophage T7 / genetics
DNA-Directed RNA Polymerases / chemistry
DNA-Directed RNA Polymerases / genetics*
Directed Molecular Evolution*
Feedback, Physiological
Genes, Viral
Nucleic Acid Conformation
RNA, Messenger / chemistry
RNA, Messenger / genetics
RNA, Viral / chemistry
RNA, Viral / genetics
Synthetic Biology
Viral Proteins / chemistry
Viral Proteins / genetics*

Substances

RNA, Messenger
RNA, Viral
Viral Proteins
RNA polymerase SP6
bacteriophage T7 RNA polymerase
DNA-Directed RNA Polymerases