ShanghaiTech University Knowledge Management System
| Cell extract-based in vitro DNA replication enables sustainable cell-free gene expression | |
| 2024-10-02 | |
| 状态 | 已发表 |
| 摘要 | Cell extract-based cell-free gene expression (CFE) systems break cellular boundaries and create emerging platforms for synthetic biology applications. However, these systems lack intrinsic DNA replication capability, thereby limiting sustained transcription and translation in a cell-free environment. Here, we introduce a LoopReX system based on E. coli CFE by integration of the DNA replication machinery, enabling iterative in vitro DNA replication and sustainable cell-free gene expression. We establish LoopReX by integrating phi29 DNA polymerase into the crude lysate-based E. coli CFE system, allowing for coupled DNA replication and protein expression. We elucidate the synergetic mechanisms between T7 RNA polymerase and phi29 DNA polymerase in facilitating in vitro DNA replication. Additionally, we optimize LoopReX reaction conditions by machine learning strategies. Finally, we demonstrate that the enhanced LoopReX system achieves highly efficient DNA replication and protein production in an iterative, scalable, and sustainable manner. We anticipate that LoopReX will significantly advance the application of CFE in synthetic biology. |
| 语种 | 英语 |
| DOI | 10.1101/2024.10.01.616023 |
| 相关网址 | 查看原文 |
| 出处 | bioRxiv |
| 收录类别 | PPRN.PPRN |
| WOS记录号 | PPRN:100744069 |
| WOS类目 | Biology |
| 文献类型 | 预印本 |
| 条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/433523 |
| 专题 | 物质科学与技术学院 物质科学与技术学院_PI研究组_凌盛杰组 物质科学与技术学院_PI研究组_李健组 物质科学与技术学院_硕士生 物质科学与技术学院_博士生 物质科学与技术学院_PI研究组_刘一凡组 |
| 通讯作者 | Ling, Shengjie; Li, Jian |
| 作者单位 | 1.ShanghaiTech Univ, Sch Phys Sci & Technol, Shanghai 201210, Peoples R China 2.ShanghaiTech Univ, State Key Lab Adv Med Mat & Devices, Shanghai 201210, Peoples R China 3.Shanghai Clin Res & Trial Ctr, Shanghai 201210, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zheng, Xiao,Gao, Wenli,Liu, Wan-Qiu,et al. Cell extract-based in vitro DNA replication enables sustainable cell-free gene expression. 2024. |
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