Cell-free gene expression
Cell-free gene expression
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DOI：
10.1038/s43586-021-00046-x
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发表时间：
2021-07-15
期刊：
影响因子：
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通讯作者：
Noireaux, Vincent
中科院分区：
文献类型：
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作者：
Garenne, David;Haines, Matthew C.;Noireaux, Vincent
无细胞基因表达（CFE）作为活细胞的替代方法出现，用于结构生物学和蛋白质组学研究的蛋白质合成和标记的特定应用。此后，CFE 被重新用作合成生物学和生物工程的多功能技术。然而，充分利用这项技术需要深入了解现有协议之外的基本工作流程。本入门指南为新从业者提供了全面、详细且可操作的 CFE 最佳实践指南，为最先进的实验室研究提供信息。我们专注于基于大肠杆菌的 CFE 系统，该系统仍然是高效 CFE 的主要平台。生产蛋白质、生物制造疗法、开发传感器和原型遗传电路说明了 CFE 的实用介绍所提供的更广泛的实用性和机会。凭借其广泛的功能和可移植性，CFE 正在成为生物技术领域强大且有利的研究工具。
Cell-free gene expression (CFE) emerged as an alternative approach to living cells for specific applications in protein synthesis and labelling for structural biology and proteomics studies. CFE has since been repurposed as a versatile technology for synthetic biology and bioengineering. However, taking full advantage of this technology requires in-depth understanding of its fundamental workflow beyond existing protocols. This Primer provides new practitioners with a comprehensive, detailed and actionable guide to best practices in CFE, to inform research in the laboratory at the state of the art. We focus on Escherichia coli-based CFE systems, which remain the primary platform for efficient CFE. Producing proteins, biomanufacturing therapeutics, developing sensors and prototyping genetic circuits illustrate the broader utility and opportunities provided by this practical introduction to CFE. With its extensive functionality and portability, CFE is becoming a powerful and enabling research tool for biotechnology.