Automation and Expansion of EMMA Assembly for Fast-Tracking Mammalian System Engineering

Joshua James, Sally Jones, Andrea Martella, Yisha Luo, David I. Fisher, Patrick Cai

Research output: Contribution to journalArticlepeer-review

Abstract

With applications from functional genomics to the production of therapeutic biologics, libraries of mammalian expression vectors have become a cornerstone of modern biological investigation and engineering. Multiple modular vector platforms facilitate the rapid design and assembly of vectors. However, such systems approach a technical bottleneck when a library of bespoke vectors is required. Utilising the flexibility and robustness of the Extensible Mammalian Modular Assembly toolkit (EMMA), we present an automated work flow for the library-scale design, assembly and verification of mammalian expression vectors. Vector design is simplified using our EMMA computer-aided design tool (EMMA-CAD), whilst the precision and speed of acoustic droplet ejection technology are applied in vector assembly. Our pipeline facilitates significant reductions in both reagent usage and researcher hands-on time when compared with manual assembly, as shown by system Q-metrics. To demonstrate automated EMMA performance, we compiled a library of 48 distinct plasmid vectors, encoding either CRISPR interference or activation modalities. Characterization of work flow parameters shows that high assembly efficiency is maintained across vectors of various sizes and design complexities. Our system also performs strongly when compared to manual assembly efficiency benchmarks. Alongside our automated pipeline, we present a straightforward strategy for integrating gRNA and Cas9 modules into the EMMA platform, enabling the design and manufacture of valuable genome editing resources.
Original languageEnglish
JournalACS Synthetic Biology
Publication statusAccepted/In press - 21 Dec 2021

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