Genome replication, synthesis, and assembly of the bacteriophage T7 in a single cell-Free reaction

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Abstract

The synthesis of living entities in the laboratory is a standing challenge that calls for innovative approaches. Using a cell-free transcriptiontranslation system as a molecular programming platform, we show that the bacteriophage T7, encoded by a 40 kbp DNA program composed of about 60 genes, can be entirely synthesized from its genomic DNA in a test tube reaction. More than a billion infectious bacteriophages T7 per milliliter of reaction are produced after a few hours of incubation. The replication of the genomic DNA occurs concurrently with phage gene expression, protein synthesis, and viral assembly. The demonstration that genome-sized viral DNA can be expressed in a test tube, recapitulating the entire chain of information processing including the replication of the DNA instructions, opens new possibilities to program and to study complex biochemical systems in vitro.

Original languageEnglish (US)
Pages (from-to)408-413
Number of pages6
JournalACS Synthetic Biology
Volume1
Issue number9
DOIs
StatePublished - Sep 21 2012

Keywords

  • Bacteriophage
  • Cell-free protein synthesis
  • Cell-free synthetic biology
  • DNA replication
  • Self-assembly

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