The DOI matches its Crossref record; 56 of the 56 references it cites were found in a registry or an index.
CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii
Open accessAbstract
Genome editing is crucial for genetic engineering of organisms for improved traits, particularly in microalgae due to the urgent necessity for the next generation biofuel production. The most advanced CRISPR/Cas9 system is simple, efficient and accurate in some organisms; however, it has proven extremely difficult in microalgae including the model alga Chlamydomonas. We solved this problem by delivering Cas9 ribonucleoproteins (RNPs) comprising the Cas9 protein and sgRNAs to avoid cytotoxicity and off-targeting associated with vector-driven expression of Cas9. We obtained CRISPR/Cas9-induced mutations at three loci including MAA7, CpSRP43 and ChlM, and targeted mutagenic efficiency was improved up to 100 fold compared to the first report of transgenic Cas9-induced mutagenesis. Interestingly, we found that unrelated vectors used for the selection purpose were predominantly integrated at the Cas9 cut site, indicative of NHEJ-mediated knock-in events. As expected with Cas9 RNPs, no off-targeting was found in one of the mutagenic screens. In conclusion, we improved the knockout efficiency by using Cas9 RNPs, which opens great opportunities not only for biological research but also industrial applications in Chlamydomonas and other microalgae. Findings of the NHEJ-mediated knock-in events will allow applications of the CRISPR/Cas9 system in microalgae, including “safe harboring” techniques shown in other organisms.
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A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998) · Nature · doi:10.1038/35888 · Cited by 15,429
The new frontier of genome engineering with CRISPR-Cas9
Doudna, J. A., & Charpentier, E. (2014) · Science · doi:10.1126/science.1258096 · Cited by 7,062
Development and Applications of CRISPR-Cas9 for Genome Engineering
Hsu, P. D., Lander, E. S., & Zhang, F. (2014) · Cell · doi:10.1016/j.cell.2014.05.010 · Cited by 5,762
SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler
Luo, R., Liu, B., Xie, Y., Li, Z., Huang, W., Yuan, J., He, G., Chen, Y., Pan, Q., Liu, Y., et al. (2012) · Gigascience · doi:10.1186/2047-217x-1-18 · Cited by 5,307
ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering
Gaj, T., Gersbach, C. A., & Barbas, C. F. (2013) · Trends in Biotechnology · doi:10.1016/j.tibtech.2013.04.004 · Cited by 3,820
The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway
Lieber, M. R. (2010) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.052308.093131 · Cited by 2,882
The Chlamydomonas Genome Reveals the Evolution of Key Animal and Plant Functions
Merchant, S. S., Prochnik, S. E., Vallon, O., Harris, E. H., Karpowicz, S. J., Witman, G. B., Terry, A., Salamov, A., Fritz-Laylin, L. K., Maréchal-Drouard, L., et al. (2007) · Science · doi:10.1126/science.1143609 · Cited by 2,691
Bae, S., Park, J., & Kim, J.-S. (2014) · Bioinformatics · doi:10.1093/bioinformatics/btu048 · Cited by 2,339
Self-citationWang, Y., Cheng, X., Shan, Q., Zhang, Y., Liu, J., Gao, C., & Qiu, J.-L. (2014) · Nature Biotechnology · doi:10.1038/nbt.2969 · Cited by 2,072
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Abdullah, B., Syed Muhammad, S. A. F., Shokravi, Z., Ismail, S., Kassim, K. A., Mahmood, A. N., & Aziz, M. M. A. (2019) · Renewable and Sustainable Energy Reviews · doi:10.1016/j.rser.2019.02.018 · Cited by 531
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Alishah Aratboni, H., Rafiei, N., Garcia-Granados, R., Alemzadeh, A., & Morones-Ramírez, J. R. (2019) · Microbial Cell Factories · doi:10.1186/s12934-019-1228-4 · Cited by 437
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Metadata
- Authors
- Shin, S.-E., Lim, J.-M., Koh, H. G., Kim, E. K., Kang, N. K., Jeon, S., Kwon, S., Shin, W.-S., Lee, B., Hwangbo, K., Kim, J., Ye, S. H., Yun, J.-Y., Seo, H., Oh, H.-M., Kim, K.-J., Kim, J.-S., Jeong, W.-J., Chang, Y. K., & Jeong, B.-R.
- Venue
- Scientific Reports
- Published
- 2016-06-13
- Volume
- 6
- Issue
- 1
- Pages
- 27810-27810
- ISSN
- 2045-2322 (ISSN-L)
- OpenAlex
- W2443008206
- Cited by
- 423
- References
- 56
- FWCI
- 19.56
- Type
- journal article
- Language
- en
- Open access
- gold
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- Shin, S.-E., Lim, J.-M., Koh, H. G., Kim, E. K., Kang, N. K., Jeon, S., Kwon, S., Shin, W.-S., Lee, B., Hwangbo, K., Kim, J., Ye, S. H., Yun, J.-Y., Seo, H., Oh, H.-M., Kim, K.-J., Kim, J.-S., Jeong, W.-J., Chang, Y. K., & Jeong, B.-R. (2016). CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii. Scientific Reports. https://doi.org/10.1038/srep27810
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