Repurposing of Anthocyanin Biosynthesis for Plant Transformation and Genome Editing

• Published in: Frontiers in genome editing Vol. 2; p. 607982
• Main Authors: He, Yubing, Zhu, Min, Wu, Junhua, Ouyang, Lejun, Wang, Rongchen, Sun, Hui, Yan, Lang, Wang, Lihao, Xu, Meilian, Zhan, Huadong, Zhao, Yunde
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 03.12.2020
• Subjects: AAC; anthocyanin; CRISPR; Genome Editing; rice; transgene-free; CRISPR; AAC; anthocyanin; rice; transgene-free
• ISSN: 2673-3439; 2673-3439
• DOI: 10.3389/fgeed.2020.607982

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.