State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Nat Commun. 2016 Aug 25;7:12617. doi: 10.1038/ncomms12617.
Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.
在难以转化的植物中编辑植物基因组在技术上具有挑战性,并且通常涉及转基因中间体,这引起了监管方面的关注。在这里,我们报告了两种简单有效的基因组编辑方法,其中植物是从瞬时表达 CRISPR/Cas9 的愈伤组织细胞再生的,该 CRISPR/Cas9 是作为 DNA 或 RNA 引入的。这种基于瞬时表达的基因组编辑系统在 T0 代产生无转基因和纯合小麦突变体方面非常高效和特异。我们展示了我们在六倍体面包小麦和四倍体硬粒小麦中编辑基因的方案,并表明我们能够生成无可检测转基因的突变体。我们的方法可能适用于其他植物物种,从而有可能加速基础和应用植物基因组工程研究。
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