Cell-Free Synthetic Biology Platform for Engineering Synthetic Biological Circuits and Systems.

Jeongwon Kim,Elisa Franco,Melissa Klocke,Siddharth Agarwal,Seungdo Choi,Dohyun Jeong,Jongmin Kim

doi:10.3390/mps2020039

Abstract

Synthetic biology integrates diverse engineering disciplines to create novel biological systems for biomedical and technological applications. The substantial growth of the synthetic biology field in the past decade is poised to transform biotechnology and medicine. To streamline design processes and facilitate debugging of complex synthetic circuits, cell-free synthetic biology approaches has reached broad research communities both in academia and industry. By recapitulating gene expression systems in vitro, cell-free expression systems offer flexibility to explore beyond the confines of living cells and allow networking of synthetic and natural systems. Here, we review the capabilities of the current cell-free platforms, focusing on nucleic acid-based molecular programs and circuit construction. We survey the recent developments including cell-free transcription–translation platforms, DNA nanostructures and circuits, and novel classes of riboregulators. The links to mathematical models and the prospects of cell-free synthetic biology platforms will also be discussed.

Highlights

Synthetic biology focuses on engineering biological circuits to manipulate biological systems and technological applications
We review the current scope of cell-free synthetic biology, focusing on synthetic circuits and systems using nucleic acid-based programs
We limit ourselves to the design and MethodIsnPrtohtoics. 2a0r1t9i,c2le, 3, 9we review the current scope of cell-free synthetic biology, focusing on syn2thofe2ti5c circuits and systems using nucleic acid-based programs

Summary

Introduction

Synthetic biology focuses on engineering biological circuits to manipulate biological systems and technological applications. Formative works in synthetic biology demonstrated the creation of simple regulatory circuits in Escherichia coli [1,2]. Cell-free synthetic biology provides a paradigm to test components and circuits in a well-controlled environment that is similar to physiological conditions [6]. We review the current scope of cell-free synthetic biology, focusing on synthetic circuits and systems using nucleic acid-based programs. 2a0r1t9i,c2le, 3, 9we review the current scope of cell-free synthetic biology, focusing on syn2thofe2ti5c circuits and systems using nucleic acid-based programs. We limit ourselves to the design and aapppplliiccaattiioonnss ooff tthheessee ssyynntthheettiicc mmoolleeccuullaarrcciirrccuuititss..RReaedaedresrasraerreerfeerfererrdedtotoothoethr eerxceexlcleenlltenretvrieevwieswfosr froercerenctendtevdeelvoeplmopemntesntins inothoethr earraeraesasofofceclell-lf-rfereeessyynnththeetitcic bbiioollooggyy ssuucchh aass cceellll--ffrreeee mmeetatabboolilcic eennggiinneeeerriinngg [[77,8,8]]. The concentration of individual components can be adjusted in the PURE system during

Methods

Methods and Protoc

RNA Regulatory Circuits for Cell-Free Synthetic Biology

Encapsulation of in Vitro Circuits toward the Synthesis of Artificial Cells

Concluding Remarks