Multiplex transcriptional characterizations across diverse bacterial species using cell‐free systems
Year: 2019
Type: article
Abstract: Cell-free expression systems enable rapid prototyping of genetic programs in vitro. However, current throughput of cell-free measurements is limited by the use of channel-limited fluorescent readouts. 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 employ this method in active cell lysates developed from ten diverse bacterial species. Interspecies analysis of transcriptional profiles from > 1,000 diverse regulatory sequences reveals functional differences in promoter activity that can be quantitatively modeled, providing a rich resource for tuning gene expression in diverse bacterial species. Finally, we examine the transcriptional capacities of dual-species hybrid lysates that can simultaneously harness gene expression properties of multiple organisms. We expect that this cell-free multiplex transcriptional measurement approach will improve genetic part prototyping in new bacterial chassis for synthetic biology.
Source: Molecular Systems Biology
Authors: Sung Sun Yim, Nathan I. Johns, Jimin Park, Antonio L.C. Gomes, Ross M. McBee, Miles Richardson, Carlotta Ronda, Sway P. Chen, David Garenne, Vincent Noireaux, Harris H. Wang
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $4,200
Funders: National Science Foundation, Alfred P. Sloan Foundation, Human Frontier Science Program, National Research Foundation, National Research Foundation of Korea, Korea Advanced Institute of Science and Technology, National Institutes of Health, Defense Advanced Research Projects Agency, U.S. Navy, Office of Naval Research, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases
Awards: G1163247334, G1220657860, G1378838858, G1544635806, G1728801193, G1760589713, A minimally invasive synthetic bio-driven approach for natural products discovery, G214439962, G2515541506, G2518502058, G2558686711, G2647030984, G2704988860, G273449253, G3328164658, G3624902447, G3892866308, G4198750475, G4252360166, G4324411131, G4400530034, Graduate Research Fellowship Program (GRFP), G5584835802, G5691479906, G5850534, CAREER: A Systems Approach to Study Horizontal Acquisition of Regulatory DNA, G59742032, G6206737223, G624019939, Towards self-reproduction of protocells and minimal cells: evolution versus engineering, G7832175178, G8132658730, G8238768891, G8876996369, Micron-scale Spatial Metagenomic Mapping of Microbial Biogeography in the Gastrointestinal Tract, G910434835, G960570965
OpenAlex ID: W2968684239