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This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.Abstract The CRISPR/Cas9 is a core technology that can result in a paradigm for breeding new varieties. This study describes in detail the sgRNA design, vector construction, and the development of a transgenic plant and its molecular analysis, and demonstrates how gene editing technology through the CRISPR/Cas9 system can be applied easily and accurately. CRISPR/Cas9 facilitates targeted gene editing through RNA-guided DNA cleavage, followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. It also allows the generation of heritabletargeted gene mutations and corrections. Here, we present detailed procedures involved in the CRISPR/Cas9 system to acquire faster, easier and more cost-efficient gene edited transgenic rice. The protocol described here establishes the strategies and steps for the selection of targets, design of sgRNA, vector construction, and analysis of the transgenic lines. The same principles can be used to customize the versatile CRISPR/Cas9 system, for application to other plant species.Keywords CRISPR/Cas9, Gene editing, sgRNA design, transgenic rice, null plant

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This study was conducted from 2010 to May 2012 to determine the volatile flavor compositions, biological activity and components of A. monanthum from different regions in Korea. The flavors of A. monanthum were extracted by SPME methods and it contained forty-two compounds that included mainly hydrocarbons and acids. The cancer cell growth inhibition activities of A. monanthum on the cancer cell (HaCaT, HepG2, HCT116, PC3) line were increased in a dose-dependent manner and the hexane fraction showed the highest antiproliferation effects. A. monanthum also showed the highest antioxidant activity. The results suggest that A. monanthum can be used as bioactive and functional materials.

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