In vitro DNA SCRaMbLE

• DOI: 10.1038/s41467-018-03743-6
• Publication Date: 2018-05-10T09:20:46Z
• Authors: Yi Wu; Rui-Ying Zhu; Leslie A. Mitchell; Lu Ma; Rui Liu; Meng Zhao; Bin Jia; Hui Xu; Yun-Xiang Li; Zu-Ming Yang; Yuan Ma; Xia Li; Hong Liu; Duo Liu; Wen-Hai Xiao; Xiao Zhou; Bing-Zhi Li; Ying-Jin Yuan; Jef D. Boeke
• Keywords: Synthetic Biology

Abstract The power of synthetic biology has enabled the expression heterologous pathways in cells, as well genome-scale synthesis projects. complexity biological networks makes rational de novo design a grand challenge. Introducing features that confer genetic flexibility is powerful strategy for downstream engineering. Here we develop an vitro method DNA library construction based on structural variation to accomplish this goal. “in SCRaMbLE system” uses Cre recombinase mixed test tube with purified encoding multiple loxPsym sites. Using β-carotene pathway designed yeast example, demonstrate top-down and bottom-up SCRaMbLE, enabling optimization biosynthetic flux via rearrangement relevant transcription units. We show our system provides straightforward way correlate phenotype genotype potentially amenable biochemical ways vivo cannot achieve.