RNA-guided Cas9 as an in vivo desired-target mutator in maize.
RNA-guided Cas9 as an in vivo desired-target mutator in maize.
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DOI：
10.1111/pbi.12739
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发表时间：
2017-12
影响因子：
13.8
通讯作者：
Xie C
中科院分区：
文献类型：
--
作者：
Li C;Liu C;Qi X;Wu Y;Fei X;Mao L;Cheng B;Li X;Xie C
RNA引导的Cas9系统是基因组编辑的通用工具。在这里，我们建立了一个RNA引导的核酸内切酶（RGEN）系统，作为玉米体内所需的目标突变体（DTM），以减少育种过程中的连锁阻力，使用LIGULELESS 1（LG 1）基因座作为概念验证。在T0代转基因植株中，突变频率为51.5%-91.2%。然后，我们将稳定表达DTM的T1植株与6个不同的受体玉米品系杂交，发现11.79%-28.71%的受试植株是由DTM效应诱导的突变体。连续F2植株的分析表明，DTM效应诱导的突变在很大程度上是可遗传的。此外，DTM产生的杂种具有显著更小的叶角，与野生型相比减少了50%以上。种植实验表明，DTM产生的玉米植株可以以显著更高的密度生长，因此具有更大的产量潜力。我们的工作表明，稳定表达的RGEN可以作为体内DTM来实施，以通过杂交和随后的回交在玉米中快速产生和传播所需的突变，从而绕过常规渐渗方法中的连锁拖拽效应。这种概念验证实验可能是采用RNA指导的Cas9基因组编辑的作物中潜在的更有效的育种策略。
The RNA‐guided Cas9 system is a versatile tool for genome editing. Here, we established a RNA‐guided endonuclease (RGEN) system as an in vivo desired‐target mutator (DTM) in maize to reduce the linkage drag during breeding procedure, using the LIGULELESS1 (LG1) locus as a proof‐of‐concept. Our system showed 51.5%–91.2% mutation frequency in T0 transgenic plants. We then crossed the T1 plants stably expressing DTM with six diverse recipient maize lines and found that 11.79%–28.71% of the plants tested were mutants induced by the DTM effect. Analysis of successive F2 plants indicated that the mutations induced by the DTM effect were largely heritable. Moreover, DTM‐generated hybrids had significantly smaller leaf angles that were reduced more than 50% when compared with that of the wild type. Planting experiments showed that DTM‐generated maize plants can be grown with significantly higher density and hence greater yield potential. Our work demonstrate that stably expressed RGEN could be implemented as an in vivo DTM to rapidly generate and spread desired mutations in maize through hybridization and subsequent backcrossing, and hence bypassing the linkage drag effect in convention introgression methodology. This proof‐of‐concept experiment can be a potentially much more efficient breeding strategy in crops employing the RNA‐guided Cas9 genome editing.
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影响因子：
5.1
作者：
REGISTER, JC;PETERSON, DJ;WILSON, HM
通讯作者：
WILSON, HM
影响因子：
56.9
作者：
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通讯作者：
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27.5
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
11.6
作者：
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通讯作者：
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