无细胞蛋白质合成：从基础研究到工程应用

后佳琦; 姜楠; 马莲菊; 卢元

doi:10.12211/2096-8280.2021-064

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无细胞蛋白质合成：从基础研究到工程应用

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- 无细胞蛋白质合成：从基础研究到工程应用
- Cell-free protein synthesis: from basic research to engineering applications
- 合成生物学 2022年3卷第3期页码：465-486
- 作者机构：
  
  1.清华大学化学工程系，北京 100084
  2.沈阳师范大学生命科学学院，辽宁沈阳 110034
- 作者简介：
  
  [ "后佳琦（1994—），女，硕士研究生。研究方向为无细胞生物合成。 E-mail：2580070932@qq.com" ]
  [ "姜楠（1995—），女，硕士研究生。研究方向为无细胞生物合成。 E-mail：1023796982@qq.com" ]
  [ "马莲菊（1969—），女，教授，硕士生导师。研究方向为资源与应用微生物学等。 E-mail：malianju@163.com" ]
  [ "卢元（1983—），男，副教授，博士生导师。研究方向为合成生物学、生物大分子工程等。 E-mail：yuanlu@tsinghua.edu.cn" ]
- 基金信息：
  
  国家自然科学基金(21878173);北京市自然科学基金(2192023);国家重点研发计划(2018YFA0901700);清华大学实验室创新基金
- DOI：10.12211/2096-8280.2021-064
  中图分类号： Q816
- 收稿：2021-06-06，
  
  修回：2021-09-21，
  
  纸质出版：2022-06-30
- 稿件说明：
移动端阅览
后佳琦，姜楠，马莲菊，卢元. 无细胞蛋白质合成：从基础研究到工程应用［J］. 合成生物学， 2022， 3（3）： 465-486

HOU Jiaqi， JIANG Nan， MA Lianju， LU Yuan. Cell-free protein synthesis： from basic research to engineering applications［J］. Synthetic Biology Journal， 2022， 3（3）： 465-486
后佳琦，姜楠，马莲菊，卢元. 无细胞蛋白质合成：从基础研究到工程应用［J］. 合成生物学， 2022， 3（3）： 465-486 DOI： 10.12211/2096-8280.2021-064.

HOU Jiaqi， JIANG Nan， MA Lianju， LU Yuan. Cell-free protein synthesis： from basic research to engineering applications［J］. Synthetic Biology Journal， 2022， 3（3）： 465-486 DOI： 10.12211/2096-8280.2021-064.

摘要

无细胞蛋白质合成是无细胞合成生物学的技术核心，亦被称为体外蛋白质翻译，是一种用于补充基于细胞的蛋白质表达的技术。无细胞蛋白质合成系统无需完整的活细胞就可以在体外受控环境中模拟整个细胞的转录和翻译过程，并允许对单个成分和反应网络进行详细深入的研究。因此，无细胞蛋白质合成作为一种平台技术，有望克服当前胞内生产系统中因为细胞膜约束带来的表达局限性，在基础科学研究和应用科学研究中具有广阔的前景。无细胞系统操作简单、便于控制，相对于体内蛋白质表达，其优势还包括其开放特性、消除对活细胞的依赖以及将所有系统物质能量集中在目标蛋白质生产上。本文首先概述了无细胞蛋白质合成系统的组成及基于不同组件类型的无细胞蛋白质合成系统的发展，包括以不同生物提取物为基础的系统以及使用重组元素的蛋白质合成体系。之后介绍了以分批反应、连续交换为代表的无细胞蛋白质合成系统的不同反应模式，阐述了无细胞在基因电路、蛋白质工程和人工“生命体系”构建中的应用和研究进展。其中，基因电路主要概述了无细胞技术在原型设计、生物传感、代谢工程三个方面的最新应用；蛋白质工程依次罗列了无细胞技术在膜蛋白、类病毒颗粒、翻译后修饰、非天然氨基酸嵌入以及蛋白质进化等方面的应用拓展；在人工“生命体系”构建中，噬菌体的合成和人工细胞的构筑开辟了新的前沿领域。最后文章分析了无细胞蛋白质合成系统在未来进一步的科学研究和工业化应用中面临的机遇和挑战。

Abstract

Cell-free protein synthesis (CFPS)

also known as

in vitro

gene expression

is a multifunctional technique used to complement cell-based protein expression

which is at the core of cell-free synthetic biology. Since the CFPS system does not require a living cell

it can simulate the entire cellular transcription and translation process

in vitro

in a controlled environment

and allows for an in-depth study of individual components and biological networks. Therefore

as a platform technology

it is expected to overcome the loopholes caused by the limitations of cell membranes in the current

in vivo

manufacturing systems

which has a broad research prospect in fundamental and applied scientific research. The cell-free operation is simple and easy to control

and its advantages over

in vivo

protein expression include its nature with open systems

eliminating the dependence on living cells and using all system energy for the production of the target proteins. This article reviews the composition of CFPS systems and their development based on different component types

including different biological extracts or purified transcription and translation components. Furthermore

different CFPS reaction patterns are introduced

inclu

ding batch and continuous exchange modes

and the research progress of CFPS systems in genetic circuits

protein engineering

and the construction of artificial life is described. Among them

the genetic circuit research progress mainly summarizes the latest applications and contributions of cell-free technology in the prototype design

biosensors

and in vitro metabolic engineering. The protein engineering research progress lists the advantages and advances of the CFPS systems for producing membrane proteins

virus-like particles

post-translational modifications

unnatural amino acid incorporation and protein evolution. In the construction of artificial "living systems"

the synthesis of bacteriophages and the construction of artificial cells have opened up a novel frontier field. Finally

the opportunities and challenges of the CFPS platforms for future scientific research and industrial applications are highlighted.

关键词

Keywords

references

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