TY - JOUR T1 - Multiplex transcriptional characterizations across diverse and hybrid bacterial cell-free expression systems JF - bioRxiv DO - 10.1101/427559 SP - 427559 AU - Sung Sun Yim AU - Nathan I. Johns AU - Jimin Park AU - Antonio L.C. Gomes AU - Ross M. McBee AU - Miles Richardson AU - Carlotta Ronda AU - Sway P. Chen AU - David Garenne AU - Vincent Noireaux AU - Harris H. Wang Y1 - 2018/01/01 UR - http://biorxiv.org/content/early/2018/09/26/427559.abstract N2 - 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. ER -