期刊
PLANT BIOTECHNOLOGY JOURNAL
卷 19, 期 10, 页码 2000-2010出版社
WILEY
DOI: 10.1111/pbi.13613
关键词
CRISPR-Cas9; genome editing; targeted gene insertion; maize; Agrobacterium transformation
CRISPR-Cas is a powerful tool for DNA double-strand break technology, with wide applications in plant genome modification. Advancements in maize transformation and vector design have greatly increased the efficiency of targeted gene insertion, allowing for potential trait improvement in a wide range of crop species.
CRISPR-Cas is a powerful DNA double-strand break technology with wide-ranging applications in plant genome modification. However, the efficiency of genome editing depends on various factors including plant genetic transformation processes and types of modifications desired. Agrobacterium infection is the preferred method of transformation and delivery of editing components into the plant cell. While this method has been successfully used to generate gene knockouts in multiple crops, precise nucleotide replacement and especially gene insertion into a pre-defined genomic location remain highly challenging. Here, we report an efficient, selectable marker-free site-specific gene insertion in maize using Agrobacterium infection. Advancements in maize transformation and new vector design enabled increase of targeted insertion frequencies by two orders of magnitude in comparison to conventional Agrobacterium-mediated delivery. Importantly, these advancements allowed not only a significant improvement of the frequency, but also of the quality of generated events. These results further enable the application of genome editing for trait product development in a wide variety of crop species amenable to Agrobacterium-mediated transformation.
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