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. 2020 Dec 17;10:22158. doi: 10.1038/s41598-020-78968-x

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Delivery efficiency of RNP and Cytotoxicity of Cas9 and ribonucleoprotein. (a) The cellular uptake efficiency of Cy3-RNP into wild-type C. reinhardtii measured by a flow cytometer according to the concentration of pVEC. The ratio of the quantum yield of wild-type C. reinhardtii according to the concentration of pVEC. After incubation of samples for one hour under dark as a dark adaptation, quantum yield was measured by AquaPen-C APA100. The ratio of the quantum yield was calculated by dividing the QY of the experimental group by the QY of the control group. The data represent the average of n = 3 replicate experiments. Standard deviation bars are shown. **Significantly different (Student’s t-test, p < 0.05). *Significantly not different (Student’s t-test, p > 0.05). (b) The recovery of algal cell on agar plate after treatment. Relative protein amount 1 represents 10ug of protein (Cas9 and alcohol dehydrogenase). 600 cells were spread and counted. The final concentration of pVEC for delivery of protein was 8uM. **Significantly different (Student’s t-test, p < 0.05). (c) The quantum yield for algal cell depends on time. Relative amount 1 of protein was used. The quantum yield was measured by AquaPen-C APA100. The data represent the average of n = 3 replicate experiments. Standard deviation bars are not shown for legibility.