DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins
DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins
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DOI：
10.1038/nbt.3389
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发表时间：
2015-11-01
影响因子：
46.9
通讯作者：
Kim, Jin-Soo
中科院分区：
文献类型：
--
作者：
Woo, Je Wook;Kim, Jungeun;Kim, Jin-Soo
编辑植物基因组而不将外源DNA引入细胞可能会减轻与转基因植物相关的监管问题。我们将纯化的Cas9蛋白和指导RNA的预组装复合物转染到拟南芥、烟草、莴苣和水稻的植物原生质体中，并以高达46%的频率在再生植物中实现了靶向诱变。靶向位点含有生殖系传播的小插入或缺失，这些插入或缺失与自然发生的遗传变异无法区分。
Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.