Precise insertion and guided editing of higher plant genomes using Cpf1 CRISPR nucleases.
Precise insertion and guided editing of higher plant genomes using Cpf1 CRISPR nucleases.
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DOI：
10.1038/s41598-017-11760-6
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发表时间：
2017-09-14
影响因子：
4.6
通讯作者：
Oufattole M
中科院分区：
文献类型：
--
作者：
Begemann MB;Gray BN;January E;Gordon GC;He Y;Liu H;Wu X;Brutnell TP;Mockler TC;Oufattole M
植物的精确基因组编辑有可能通过内源途径的靶向工程或引入新性状来重塑全球农业。为了开发一种基于CRISPR核酸酶的平台，以实现更高效率的精确基因插入或替换，我们筛选了来自新杀弗朗西斯菌和毛螺菌科细菌ND 2006的Cpf1核酸酶，以了解它们通过同源性定向修复诱导靶向基因插入的能力。两种核酸酶，在指导RNA和修复DNA模板的存在下，侧翼同源DNA片段的目标位点，被证明产生精确的基因插入以及插入缺失突变的目标位点在水稻基因组中。这些Cpf1核酸酶的靶向插入频率高达8%，高于大多数其他基因组编辑核酸酶，表明其有效的酶化学。Cpf1基因组编辑技术的进一步改进和广泛采用有可能对植物生物技术产生巨大影响。
Precise genome editing of plants has the potential to reshape global agriculture through the targeted engineering of endogenous pathways or the introduction of new traits. To develop a CRISPR nuclease-based platform that would enable higher efficiencies of precise gene insertion or replacement, we screened the Cpf1 nucleases from Francisella novicida and Lachnospiraceae bacterium ND2006 for their capability to induce targeted gene insertion via homology directed repair. Both nucleases, in the presence of a guide RNA and repairing DNA template flanked by homology DNA fragments to the target site, were demonstrated to generate precise gene insertions as well as indel mutations at the target site in the rice genome. The frequency of targeted insertion for these Cpf1 nucleases, up to 8%, is higher than most other genome editing nucleases, indicative of its effective enzymatic chemistry. Further refinements and broad adoption of the Cpf1 genome editing technology have the potential to make a dramatic impact on plant biotechnology.
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