He Yubing, Zhu Min, Wu Junhua, Ouyang Lejun, Wang Rongchen, Sun Hui, Yan Lang, Wang Lihao, Xu Meilian, Zhan Huadong, Zhao Yunde
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, China.
National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China.
Front Genome Ed. 2020 Dec 3;2:607982. doi: 10.3389/fgeed.2020.607982. eCollection 2020.
CRISPR/Cas9 gene editing technology has been very effective in editing genes in many plant species including rice. Here we further improve the current CRISPR/Cas9 gene editing technology in both efficiency and time needed for isolation of transgene-free and target gene-edited plants. We coupled the CRISPR/Cas9 cassette with a unit that activates anthocyanin biosynthesis, providing a visible marker for detecting the presence of transgenes. The anthocyanin-marker assisted CRISPR (AAC) technology enables us to identify transgenic events even at calli stage, to select transformants with elevated expression, and to identify transgene-free plants in the field. We used the AAC technology to edit and and successfully generated many transgene-free and target gene-edited plants at T1 generation. The AAC technology greatly reduced the labor, time, and costs needed for editing target genes in rice.
CRISPR/Cas9基因编辑技术在包括水稻在内的许多植物物种的基因编辑中都非常有效。在此，我们进一步在效率以及分离无转基因和目标基因编辑植物所需的时间方面改进了当前的CRISPR/Cas9基因编辑技术。我们将CRISPR/Cas9盒与一个激活花青素生物合成的单元相结合，提供了一个用于检测转基因存在的可见标记。花青素标记辅助CRISPR（AAC）技术使我们能够即使在愈伤组织阶段也能鉴定转基因事件，选择表达升高的转化体，并在田间鉴定无转基因植物。我们使用AAC技术编辑了 和 ，并在T1代成功产生了许多无转基因和目标基因编辑的植物。AAC技术大大减少了水稻中编辑目标基因所需的劳动力、时间和成本。
