RoboMoClo: A Robotics-Assisted Modular Cloning Framework for Multiple Gene Assembly in Biofoundry

• DOI: 10.1021/acssynbio.1c00628
• Publication Date: 2022-02-15T20:27:52Z
• Authors: Dong Hun Kang; Sung Cheon Ko; Yu Been Heo; Hyun Jeong Lee; Han Min Woo
• Keywords: Synthetic Biology; Corynebacterium glutamicum; Genome Engineering; Cloning (programming); Terminator (solar)

Efficient and versatile DNA assembly frameworks have had an impact on promoting synthetic biology to build complex biological systems. To accelerate system development, laboratory automation (or biofoundry) provides opportunity construct organisms assemblies via computer-aided design. However, a modular cloning (MoClo) for multiple limits the biofoundry workflow in terms of simplicity feasibility by preparing number materials such as destination vectors prior process. Herein, we propose robot-assisted MoClo (RoboMoClo) project with gene expressions at biofoundry. The architecture RoboMoClo framework hybrid strategy hierarchical iterative assembly, fewer compared other An industrial bacterium, Corynebacterium glutamicum, was used model host RoboMoClo. After building biopart library (promoter terminator; level 0) evaluating its features (level 1), various transcriptional directions 2) were studied using vectors. Among constructs, convergent exhibited potential interference through collision RNA polymerases. study design experiment-guided lycopene biosynthesis C. glutamicum (levels 1, 2, 3), biofoundry-assisted demonstrated proof-of-concept constructing sub-pathway units pathway 3) glutamicum. improved toolkit application.