Methodologies for preparation of prokaryotic extracts for cell-free expression systems
Abstract
Cell-free systems that mimic essential cell functions, such as gene expression, have dramatically expanded in recent years, both in terms of applications and widespread adoption. Here we provide a review of cell-extract methods, with a specific focus on prokaryotic systems. Firstly, we describe the diversity of Escherichia coli genetic strains available and their corresponding utility. We then trace the history of cell-extract methodology over the past 20 years, showing key improvements that lower the entry level for new researchers. Next, we survey the rise of new prokaryotic cell-free systems, with associated methods, and the opportunities provided. Finally, we use this historical perspective to comment on the role of methodology improvements and highlight where further improvements may be possible.
Additional Information
© 2020 Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. Peer review under responsibility of KeAi Communications Co., Ltd. Received 2 May 2020, Revised 22 July 2020, Accepted 23 July 2020, Available online 30 July 2020. We thank Richard M. Murray for review of earlier versions of the manuscript and Casey B. Bernhards, Marilyn S. Lee, Caitlin E. Sharpes, Patricia E. Buckley, and Nathan D. Mcdonald for helpful edits and feedback. We acknowledge our funding sources: the US Office of the Secretary of Defense Applied Research for the Advancement of S&T Priorities program (SDC, AEM, MWL), DARPA SBIR to Synvitrobio, Inc. dba Tierra Biosciences (ACC, ZSS), contract No: W911NF-16-P-0003, and a Caltech Grubstake Grant (ACC, ZSS). CRediT authorship contribution statement: Stephanie D. Cole: Data curation, Writing - original draft. Aleksandr E. Miklos: Data curation, Writing - original draft. Abel C. Chiao: Data curation, Writing - original draft. Zachary Z. Sun: Data curation, Writing - original draft. Matthew W. Lux: Data curation, Writing - original draft. Declaration of competing interest: ZZS and ACC have ownership in Synvitrobio, Inc. dba Tierra Biosciences, a company commercializing the applications of cell-free systems.Additional details
- Eprint ID
- 104994
- Resolver ID
- CaltechAUTHORS:20200818-102447322
- Office of the Secretary of Defense
- W911NF-16-P-0003
- Defense Advanced Research Projects Agency (DARPA)
- Caltech
- Created
-
2020-08-18Created from EPrint's datestamp field
- Updated
-
2021-11-16Created from EPrint's last_modified field