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Methodologies for preparation of prokaryotic extracts for cell-free expression systems

Cole, Stephanie D. and Miklos, Aleksandr E. and Chiao, Abel C. and Sun, Zachary Z. and Lux, Matthew W. (2020) Methodologies for preparation of prokaryotic extracts for cell-free expression systems. Synthetic and Systems Biotechnology, 5 (4). pp. 252-267. ISSN 2405-805X. https://resolver.caltech.edu/CaltechAUTHORS:20200818-102447322

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.synbio.2020.07.006DOIArticle
ORCID:
AuthorORCID
Miklos, Aleksandr E.0000-0001-6375-2304
Chiao, Abel C.0000-0002-2909-7972
Sun, Zachary Z.0000-0002-9425-2924
Lux, Matthew W.0000-0002-2773-742X
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.
Funders:
Funding AgencyGrant Number
Office of the Secretary of DefenseUNSPECIFIED
Defense Advanced Research Projects Agency (DARPA)W911NF-16-P-0003
CaltechUNSPECIFIED
Subject Keywords:Cell-free systems; Cell-free expression; Cell-free extract; Synthetic biology; Methods
Issue or Number:4
Record Number:CaltechAUTHORS:20200818-102447322
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200818-102447322
Official Citation:Stephanie D. Cole, Aleksandr E. Miklos, Abel C. Chiao, Zachary Z. Sun, Matthew W. Lux, Methodologies for preparation of prokaryotic extracts for cell-free expression systems, Synthetic and Systems Biotechnology, Volume 5, Issue 4, 2020, Pages 252-267, ISSN 2405-805X, https://doi.org/10.1016/j.synbio.2020.07.006. (http://www.sciencedirect.com/science/article/pii/S2405805X2030051X)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:104994
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:18 Aug 2020 17:47
Last Modified:18 Aug 2020 17:47

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