Open AccessAutomated Continuous Evolution of Proteins in Vivo
Author(s) -
Ziwei Zhong,
Brandon G. Wong,
Arjun Ravikumar,
Garri A. Arzumanyan,
Ahmad S. Khalil,
Chang C. Liu
Publication year - 2020
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.0c00135
Subject(s) - directed evolution , directed molecular evolution , scalability , protein engineering , experimental evolution , synthetic biology , selection (genetic algorithm) , adaptation (eye) , computational biology , mutagenesis , computer science , gene , biology , mutation , genetics , enzyme , artificial intelligence , mutant , biochemistry , database , neuroscience
We present automated continuous evolution (ACE), a platform for the hands-free directed evolution of biomolecules. ACE pairs OrthoRep, a genetic system for continuous targeted mutagenesis of user-selected genes in vivo , with eVOLVER, a scalable and automated continuous culture device for precise, multiparameter regulation of growth conditions. By implementing real-time feedback-controlled tuning of selection stringency with eVOLVER, genes of interest encoded on OrthoRep autonomously traversed multimutation adaptive pathways to reach desired functions, including drug resistance and improved enzyme activity. The durability, scalability, and speed of biomolecular evolution with ACE should be broadly applicable to protein engineering as well as prospective studies on how selection parameters and schedules shape adaptation.