Nuclear and chloroplast genome engineering of a productive non-model alga Desmodesmus armatus: Insights into unusual and selective acquisition mechanisms for foreign DNA

Damien Douchi, Megan Mosey, David Astling, Eric Knoshaug, Ambarish Nag, John McGowen, Lieve Laurens

Research output: Contribution to journalArticlepeer-review

11 Scopus Citations

Abstract

Despite the tremendous potential of algae to contribute to a future bioeconomy, there are practical and theoretical limitations to how well naturally sourced species and strains can perform in an outdoor setting. The application of biotechnology to modulate and engineer algae metabolism or to increase performance, resilience, or produce novel compounds, offers opportunities to overcome some of the major commercialization barriers. There are numerous approaches reported in the literature having variable success on genetic engineering of algae with non-model algae often presenting unique challenges to genetic engineering. We report here on successful nuclear and chloroplast genomic integration of selection marker resistance in the non-model alga Desmodesmus armatus. Nuclear transformation was accomplished using both electroporation and Agrobacterium-mediated approaches. However, in all surviving transformants, DNA integration was accompanied by excision and/or rearrangement of the gene of interest and fluorescence reporter coding sequences. Similarly, chloroplast transformation was successfully accomplished using a biolistic DNA delivery method. For these transformants, we also observed off-target mutations in the chloroplast genome, not previously observed in other, more routinely used, algae species. Finally, we present insights into potential mechanisms for these observed truncations, rearrangements, and mutations in D. armatus.

Original languageAmerican English
Article number102152
Number of pages12
JournalAlgal Research
Volume53
DOIs
StatePublished - 2021

Bibliographical note

Publisher Copyright:
© 2020

NREL Publication Number

  • NREL/JA-2700-77862

Keywords

  • Algae
  • Biolistics
  • Biorefinery
  • Genetic engineering
  • Transcriptomics
  • Transformation

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