Multiplex transcriptional characterizations across diverse and hybrid bacterial cell-free expression systems

Yim, S. S.; Johns, N. I.; Park, J.; Gomes, A. L. C.; McBee, R. M.; Richardson, M.; Ronda, C.; Chen, S. P.; Garenne, D.; Noireaux, V.; Wang, H. H.

doi:10.1101/427559

bioRxiv · 10.1101/427559

Multiplex transcriptional characterizations across diverse and hybrid bacterial cell-free expression systems

Yim, S. S.·Johns, N. I.·Park, J.·Gomes, A. L. C.·McBee, R. M.·Richardson, M.·Ronda, C.·Chen, S. P.·Garenne, D.·Noireaux, V.·Wang, H. H.

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Abstract

Cell-free expression systems enable rapid prototyping of genetic programs in vitro. However, current throughput of cell-free measurements is often limited by the use of single-channel reporter assays. Here, we describe DNA Regulatory element Analysis by cell-Free Transcription and Sequencing (DRAFTS), a rapid and robust in vitro approach for multiplexed measurement of transcriptional activities from thousands of regulatory sequences in a single reaction. We employed this method in active cell lysates developed from ten diverse bacterial species. Interspecies analysis of transcriptional profiles from >1,000 diverse regulatory sequences revealed functional differences in gene expression that could be predictively modeled. Finally, we constructed and examined the transcriptional capacities of dual-species \"hybrid\" cell lysates that can simultaneously harness gene expression properties of multiple organisms. We expect that this cell-free multiplex transcriptional measurement approach will improve genetic circuit prototyping in new bacterial chassis for synthetic biology.

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Yim, S. S., Johns, N. I., Park, J., Gomes, A. L. C., McBee, R. M., Richardson, M., Ronda, C., Chen, S. P., Garenne, D., Noireaux, V., Wang, H. H.. 2018-09-26. Multiplex transcriptional characterizations across diverse and hybrid bacterial cell-free expression systems. https://doi.org/10.1101/427559

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