The bottom-up construction of biological entities from genetic information provides a broad range of opportunities to better understand fundamental processes within living cells, as well as holding great promise for the development of novel biomedical applications. Cell-free transcription-translation (TXTL) systems have become suitable platforms to tackle such topics because they recapitulate the process of gene expression. TXTL systems have advanced to where the in vitro construction of viable, complex, self-assembling deoxyribonucleic acid-programmed biological entities is now possible. Previously, we demonstrated the cell-free synthesis of three bacteriophages from their genomes: MS2, φX174, T7. In this work, we present the complete synthesis of the phage T4 from its 169-kbp genome in one-pot TXTL reactions. This achievement, for one of the largest coliphages, demonstrates the integration of complex gene regulation, metabolism and self-assembly, and brings the bottom-up synthesis of biological systems to a new level.
Bibliographical noteFunding Information:
The Office of Naval Research [N00014-13-1-0074]; the Human Frontier Science Program [RGP0037/2015]. Noireaux laboratory receives research funds from Arbor Biosciences, a distributor of myTXTL cell-free protein expression kit.
© 2018 The Author(s) 2018. Published by Oxford University Press.
- cell-free synthetic biology
- cell-free transcription/translation