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Base Editing for Reprogramming Cyanobacterium Synechococcus elongatus

Shu-Yan Wang, Xin Li, Shu-Guang Wang, View ORCID ProfilePeng-Fei Xia
doi: https://doi.org/10.1101/2022.09.05.506134
Shu-Yan Wang
1School of Environmental Science and Engineering, Shandong University, Qingdao 266237, China
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Xin Li
1School of Environmental Science and Engineering, Shandong University, Qingdao 266237, China
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Shu-Guang Wang
1School of Environmental Science and Engineering, Shandong University, Qingdao 266237, China
2Sino-French Research Institute for Ecology and Environment, Shandong University, Qingdao 266237, China
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Peng-Fei Xia
1School of Environmental Science and Engineering, Shandong University, Qingdao 266237, China
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  • ORCID record for Peng-Fei Xia
  • For correspondence: pfxia@sdu.edu.cn
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Abstract

Global climate change demands carbon-negative innovations to reduce the concentration of atmospheric carbon dioxide (CO2). Cyanobacteria can fix CO2 from the atmosphere and can be genetically reprogrammed for the production of biofuels, chemicals and food products, making an ideal microbial chassis for carbon-negative biotechnology. However, the progress seems to be slowed down due to the lagging-behind synthetic biology toolkits, especially the CRISPR-Cas-based genome-editing tools. As such, we developed a base-editing tool based on the CRISPR-Cas system and deamination for cyanobacterium Synechococcus elongatus. We achieved efficient and precise genome editing at a single-nucleotide resolution, and identified the pure population of edited cells at the first round of selection without extra segregation. By using the base-editing tool, we successfully manipulated the glycogen metabolic pathway via the introduction of premature STOP codons to inactivate the corresponding genes. We demonstrated multiplex base editing by editing two genes at once, obtaining a nearly two-fold increase in the glycogen content. We present here the first report of base editing in the phylum of cyanobacteria, and a paradigm for applying CRISPR-Cas systems in bacteria. We believe that this work will accelerate the synthetic biology of cyanobacteria and drive more innovations to alleviate global climate change.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted September 06, 2022.
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Base Editing for Reprogramming Cyanobacterium Synechococcus elongatus
Shu-Yan Wang, Xin Li, Shu-Guang Wang, Peng-Fei Xia
bioRxiv 2022.09.05.506134; doi: https://doi.org/10.1101/2022.09.05.506134
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Base Editing for Reprogramming Cyanobacterium Synechococcus elongatus
Shu-Yan Wang, Xin Li, Shu-Guang Wang, Peng-Fei Xia
bioRxiv 2022.09.05.506134; doi: https://doi.org/10.1101/2022.09.05.506134

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