Abstract
Non-model microorganisms often possess complex phenotypes that could be important for the future of biofuel and chemical production. They have received significant interest the last several years, but advancement is still slow due to the lack of a robust genetic toolbox in most organisms. Typically, “domestication” of a new non-model microorganism has been done on an ad hoc basis, and historically, it can take years to develop transformation and basic genetic tools. Here, we review the barriers and solutions to rapid development of genetic transformation tools in new hosts, with a major focus on Restriction-Modification systems, which are a well-known and significant barrier to efficient transformation. We further explore the tools and approaches used for efficient gene deletion, DNA insertion, and heterologous gene expression. Finally, more advanced and high-throughput tools are now being developed in diverse non-model microbes, paving the way for rapid and multiplexed genome engineering for biotechnology.
| Original language | English |
|---|---|
| Article number | 30 |
| Journal | Biotechnology for Biofuels |
| Volume | 14 |
| Issue number | 1 |
| DOIs | |
| State | Published - Dec 2021 |
Funding
Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the U.S. DOE under contract DE-AC05-00OR22725. This manuscript has been authored by UT-Battelle, LLC, under Contract No. DE-AC0500OR22725 with the U.S. Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for the United States Government purposes. The Department of Energy will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan ( http://energy.gov/downloads/doe-public-access-plan ) . This work was in part supported by the Center for Bioenergy Innovation, U.S. DOE Bioenergy Research Center, supported by the Office of Biological and Environmental Research in the DOE Office of Science. This work was in part supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research, Genomic Science Program under Award Number DE-SC0019401. Funding was provided in part by the U.S. Department of Energy Office of Energy Efficiency and Renewable Energy the Bioenergy Technologies Office via the Agile BioFoundry project. The funding bodies had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.
| Funders | Funder number |
|---|---|
| Oak Ridge National Laboratory | |
| U.S. Department of Energy | DE-AC05-00OR22725 |
| U.S. Department of Energy | |
| Office of Science | |
| Biological and Environmental Research | |
| U.S. Department of Energy | |
| U.S. Department of Energy | |
| Center for Bioenergy Innovation | |
| U.S. Department of Energy Office of Energy Efficiency and Renewable Energy the Bioenergy Technologies Office | |
| Office of Science | DE-SC0019401 |
Keywords
- Genetic tools
- Genetics
- Metabolic engineering
- Non-model microbes
- Synthetic biology
- Transformation
