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  In vitro self-replication and multicistronic expression of large synthetic genomes

Libicher, K., Hornberger, R., Heymann, M., & Mutschler, H. (2020). In vitro self-replication and multicistronic expression of large synthetic genomes. Nature Communications, 11: 904. doi:10.1038/s41467-020-14694-2.

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s41467-020-14694-2.pdf (Publisher version), 2MB
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 Creators:
Libicher, K.1, Author              
Hornberger, R.1, Author              
Heymann, M.2, Author
Mutschler, H.1, Author              
Affiliations:
1Mutschler, Hannes / Biomimetic Systems, Max Planck Institute of Biochemistry, Max Planck Society, ou_2466697              
2External Organizations, ou_persistent22              

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Free keywords: DNA; DNA replication; Synthetic biology
 Abstract: The generation of a chemical system capable of replication and evolution is a key objective of synthetic biology. This could be achieved by in vitro reconstitution of a minimal self-sustaining central dogma consisting of DNA replication, transcription and translation. Here, we present an in vitro translation system, which enables self-encoded replication and expression of large DNA genomes under well-defined, cell-free conditions. In particular, we demonstrate self-replication of a multipartite genome of more than 116 kb encompassing the full set of Escherichia coli translation factors, all three ribosomal RNAs, an energy regeneration system, as well as RNA and DNA polymerases. Parallel to DNA replication, our system enables synthesis of at least 30 encoded translation factors, half of which are expressed in amounts equal to or greater than their respective input levels. Our optimized cell-free expression platform could provide a chassis for the generation of a partially self-replicating in vitro translation system.

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 Dates: 2020
 Publication Status: Published online
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 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41467-020-14694-2
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Title: Nature Communications
Source Genre: Journal
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Pages: - Volume / Issue: 11 Sequence Number: 904 Start / End Page: - Identifier: ISSN: 2041-1723