Farzadfard Fahim, Gharaei Nava, Citorik Robert J, Lu Timothy K
Synthetic Biology Group, Research Laboratory of Electronics, Department of Electrical Engineering & Computer Science and Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; MIT Synthetic Biology Center, 500 Technology Square, Cambridge, MA 02139, USA; MIT Microbiology Graduate Program, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
MCO Graduate Program, Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
Cell Syst. 2021 Sep 22;12(9):860-872.e5. doi: 10.1016/j.cels.2021.07.001. Epub 2021 Aug 5.
The ability to efficiently and dynamically change information stored in genomes would enable powerful strategies for studying cell biology and controlling cellular phenotypes. Current recombineering-mediated DNA writing platforms in bacteria are limited to specific laboratory conditions, often suffer from suboptimal editing efficiencies, and are not suitable for in situ applications. To overcome these limitations, we engineered a retroelement-mediated DNA writing system that enables efficient and precise editing of bacterial genomes without the requirement for target-specific elements or selection. We demonstrate that this DNA writing platform enables a broad range of applications, including efficient, scarless, and cis-element-independent editing of targeted microbial genomes within complex communities, the high-throughput mapping of spatial information and cellular interactions into DNA memory, and the continuous evolution of cellular traits.
高效且动态地改变基因组中存储信息的能力,将为研究细胞生物学和控制细胞表型提供强大的策略。目前细菌中基于重组工程的DNA写入平台仅限于特定的实验室条件,编辑效率往往欠佳,且不适用于原位应用。为克服这些限制,我们设计了一种逆转录元件介导的DNA写入系统,该系统能够高效、精确地编辑细菌基因组,而无需靶标特异性元件或筛选。我们证明,这个DNA写入平台具有广泛的应用,包括对复杂群落中靶向微生物基因组进行高效、无痕且不依赖顺式元件的编辑,将空间信息和细胞相互作用高通量映射到DNA记忆中,以及细胞性状的持续进化。