AUTHOR=He Yubing , Zhu Min , Wu Junhua , Ouyang Lejun , Wang Rongchen , Sun Hui , Yan Lang , Wang Lihao , Xu Meilian , Zhan Huadong , Zhao Yunde 

TITLE=Repurposing of Anthocyanin Biosynthesis for Plant Transformation and Genome Editing

JOURNAL=Frontiers in Genome Editing

VOLUME=2

YEAR=2020

URL=https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2020.607982

DOI=10.3389/fgeed.2020.607982

ISSN=2673-3439

ABSTRACT=<p>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 <italic>Cas9</italic> expression, and to identify transgene-free plants in the field. We used the AAC technology to edit <italic>LAZY1</italic> and <italic>G1</italic> 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.</p>