DNA-free two-gene knockout in Chlamydomonas reinhardtii via CRISPR-Cas9 ribonucleoproteins

Scientific Reports

Volumn 6, Issue , 2016, Pages

  Baek, Kwangryul ^a   Kim, Duk Hyoung ^a   Jeong, Jooyeon ^a   Sim, Sang Jun ^b   Melis, Anastasios ^c   Kim, Jin Soo ^d,e   Jin, Eonseon ^a   Bae, Sangsu ^a  

^a Hanyang University   (South Korea)

^b Korea University   (South Korea)

^c UNIVERSITY OF CALIFORNIA   (United States)

^d Institute for Basic Science   (South Korea)

^e Seoul National University   (South Korea)

Author keywords

[No Author keywords available]

Indexed keywords

ALGAL PROTEIN; RIBONUCLEOPROTEIN; ZEAXANTHIN;

CHLAMYDOMONAS REINHARDTII; CRISPR CAS SYSTEM; GENE KNOCKOUT; GENETICS; METABOLIC ENGINEERING; METABOLISM; PHOTOSYNTHESIS; PROCEDURES;

ALGAL PROTEINS; CHLAMYDOMONAS REINHARDTII; CRISPR-CAS SYSTEMS; GENE KNOCKOUT TECHNIQUES; METABOLIC ENGINEERING; PHOTOSYNTHESIS; RIBONUCLEOPROTEINS; ZEAXANTHINS;

EID: 84979939297     PISSN: None     EISSN: 20452322     Source Type: Journal    
DOI: 10.1038/srep30620     Document Type: Article

References (27)

1. Wijffels, R. H. & Barbosa, M. J. An outlook on microalgal biofuels. Science 329, 796-799 (2010).
2. Koller, M., Muhr, A. & Braunegg, G. Microalgae as versatile cellular factories for valued products. Algal Res. 6, 52-63 (2014).
3. Sizova, I., Greiner, A., Awasthi, M., Kateriya, S. & Hegemann, P. Nuclear gene targeting in Chlamydomonas using engineered zincfinger nucleases. Plant J. 73, 873-882 (2013).
4. Daboussi, F. et al. Genome engineering empowers the diatom Phaeodactylum tricornutum for biotechnology. Nat Commun. 5, (2014).
5. Gao, H. et al. TALE activation of endogenous genes in Chlamydomonas reinhardtii. Algal Res. 5, 52-60 (2014).
6. Jiang, W., Brueggeman, A. J., Horken, K. M., Plucinak, T. M. & Weeks, D. P. Successful transient expression of Cas9 and single guide RNA genes in Chlamydomonas reinhardtii. Eukaryot Cell 13, 1465-1469 (2014).
7. Kim, H. & Kim, J.-S. A guide to genome engineering with programmable nucleases. Nat Rev Genet. 15, 321-334 (2014).
8. Cho, S. W., Lee, J., Carroll, D., Kim, J.-S. & Lee, J. Heritable gene knockout in Caenorhabditis elegans by direct injection of Cas9-sgRNA ribonucleoproteins. Genetics 195, 1177-1180 (2013).
9. Kim, S., Kim, D., Cho, S. W., Kim, J. & Kim, J.-S. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 24, 1012-1019 (2014).
10. Zuris, J. A. et al. Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol 33, 73-80 (2015).
11. Woo, J. W. et al. DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Nat Biotechnol. 33, 1162-1164 (2015).
12. Subburaj, S. et al. Site-directed mutagenesis in Petunia hybrida protoplast system using direct delivery of purified recombinant Cas9 ribonucleoproteins. Plant Cell Rep. 35, 1535-1544 (2016).
13. Kanchiswamy, C. N., Malnoy, M., Velasco, R., Kim, J.-S. & Viola, R. Non-GMO genetically edited crop plants. Trends Biotechnol. 33, 489-491 (2015).
14. Merchant, S. S. et al. The Chlamydomonas genome reveals the evolution of key animal and plant functions. Science 318, 245-250 (2007).
15. Kirst, H., Garc-Cerd, J. G., Zurbriggen, A. & Melis, A. Assembly of the light-harvesting chlorophyll antenna in the green alga Chlamydomonas reinhardtii requires expression of the TLA2-CpFTSY gene. Plant Physiol 158, 930-945 (2012).
16. Melis, A. Solar energy conversion efficiencies in photosynthesis: minimizing the chlorophyll antennae to maximize efficiency. Plant Sci. 177, 272-280 (2009).
17. Park, J., Bae, S. & Kim, J.-S. Cas-Designer: A web-based tool for choice of CRISPR-Cas9 target sites. Bioinformatics 31, 4014-4016 (2015).
18. Bae, S., Kweon, J., Kim, H. S. & Kim, J.-S. Microhomology-based choice of Cas9 nuclease target sites. Nat Methods 11, 705-706 (2014).
19. Kindle, K. L. High-frequency nuclear transformation of Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 87, 1228-1232 (1990).
20. Bernstein, P. S. et al. Lutein, zeaxanthin, and meso-zeaxanthin: The basic and clinical science underlying carotenoid-based nutritional interventions against ocular disease. Prog Retin Eye Res. 50, 34-66 (2016).
21. Niyogi, K. K., Bjorkman, O. & Grossman, A. R. Chlamydomonas xanthophyll cycle mutants identified by video imaging of chlorophyll fluorescence quenching. Plant Cell 9, 1369-1380 (1997).
22. Jin, E., Feth, B. & Melis, A. A mutant of the green alga Dunaliella salina constitutively accumulates zeaxanthin under all growth conditions. Biotechnol Bioeng 81, 115-124 (2003).
23. Bae, S., Park, J. & Kim, J.-S. Cas-OFFinder: A fast and versatile algorithm that searches for potential off-Target sites of Cas9 RNAguided endonucleases. Bioinformatics 30, 1473-1475 (2014).
24. Kim, D. et al. Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-Target effects in human cells. Nat Methods 12, 237-243 (2015).
25. Baek, K., Lee, Y., Nam, O., Park, S., Sim, S. J. & Jin, E. Introducing Dunaliella LIP promoter containing light-inducible motifs improves transgenic expression in Chlamydomonas reinhardtii. Biotechnology Journal 11, 384-392 (2016).
26. Arnon, D. I. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24, 1 (1949).
27. Melis, A., Spangfort, M. & Andersson, B. Light-Absorption and electron-Transport balance between photosystem II and photosystem I in spinach chloroplasts. Photochem Photobiol 45, 129-136 (1987).