Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 115, Issue 3, Pages 719-728Publisher
WILEY
DOI: 10.1002/bit.26499
Keywords
algal biotechnology; Chlamydomonas reinhardtii; CRISPR-Cas9; lutein; zeaxanthin
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Funding
- Ministry of Science, ICT & Future Planning, Korea [KCRC-2014M1A8A1049273]
- Ministry of Oceans and Fisheries, Korea [20090267]
- Plant Molecular Breeding Center, Korea [PJ01119201]
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Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but the potential of this alga for carotenoid production has not yet been evaluated. In this study, we selected the C. reinhardtii CC-4349 strain as the best candidate among seven laboratory strains tested for carotenoid production. A knock-out mutant of the zeaxanthin epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins in the CC-4349 strain had a significantly higher zeaxanthin content (56-fold) and productivity (47-fold) than the wild type without the reduction in lutein level. Furthermore, we produced eggs fortified with lutein (2-fold) and zeaxanthin (2.2-fold) by feeding hens a diet containing the mutant. Our results clearly demonstrate the possibility of cost-effective commercial use of microalgal mutants induced by DNA-free CRISPR-Cas9 ribonucleoproteins in algal biotechnology for the production of high-value products.
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