Open AccessMultiplex transcriptional characterizations across diverse bacterial species using cell‐free systems
Author(s) -
Yim Sung Sun,
Johns Nathan I,
Park Jimin,
Gomes Antonio LC,
McBee Ross M,
Richardson Miles,
Ronda Carlotta,
Chen Sway P,
Garenne David,
Noireaux Vincent,
Wang Harris H
Publication year - 2019
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20198875
Subject(s) - biology , synthetic biology , computational biology , multiplex , transcriptional regulation , regulation of gene expression , gene expression , gene , crispr , genetics
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.