High efficacy full allelic CRISPR/Cas9 gene editing in tetraploid potato
High efficacy full allelic CRISPR/Cas9 gene editing in tetraploid potato
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DOI：
10.1038/s41598-019-54126-w
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发表时间：
2019-11-27
影响因子：
4.6
通讯作者：
Petersen, Bent Larsen
中科院分区：
文献类型：
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作者：
Johansen, Ida Elisabeth;Liu, Ying;Petersen, Bent Larsen
利用内源马铃薯U6启动子可显著提高四倍体马铃薯CRISPR/Cas9的编辑效率。通过用内源马铃薯U6启动子替换拟南芥U6启动子来驱动CRISPR组分的表达，大大提高了颗粒结合淀粉合成酶基因在原生质体水平上的编辑效率。这在工厂水平上转化为35%的完全等位基因编辑。Indel检测扩增分析是一种快速评估马铃薯等复杂基因组中基因编辑的有效工具。总而言之，这需要显著减少繁重的细胞培养、植物外再生和筛选具有高复杂性基因组的植物的程序。
CRISPR/Cas9 editing efficacies in tetraploid potato were highly improved through the use of endogenous potato U6 promoters. Highly increased editing efficiencies in the Granular Bound Starch Synthase gene at the protoplast level were obtained by replacement of the Arabidopsis U6 promotor, driving expression of the CRISPR component, with endogenous potato U6 promotors. This translated at the ex-plant level into 35% full allelic gene editing. Indel Detection Amplicon Analysis was established as an efficient tool for fast assessment of gene editing in complex genomes, such as potato. Together, this warrants significant reduction of laborious cell culturing, ex-plant regeneration and screening procedures of plants with high complexity genomes.