一种使用核糖核蛋白方法的简单高效的CRISPR/Cas9系统用于…… （原文结尾不完整）
A Simple and Efficient CRISPR/Cas9 System Using a Ribonucleoprotein Method for .
作者信息
Liu Jianyu, Cui Haiyang, Wang Ruijuan, Xu Zhen, Yu Hailong, Song Chunyan, Lu Huan, Li Qiaozhen, Xing Danrun, Tan Qi, Sun Weiming, Zou Gen, Shang Xiaodong
机构信息
National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China.
College of Marine Resources and Environment, Hebei Normal University of Science & Technology, Qinghuangdao 066004, China.
出版信息
J Fungi (Basel). 2022 Sep 23;8(10):1000. doi: 10.3390/jof8101000.
CRISPR/Cas9 systems were established in some edible fungi based on in vivo expressed Cas9 and guide RNA. Compared with those systems, the in vitro assembled Cas9 and sgRNA ribonucleoprotein complexes (RNPs) have more advantages, but only a few examples were reported, and the editing efficiency is relatively low. In this study, we developed and optimized a CRISPR/Cas9 genome-editing method based on in vitro assembled ribonucleoprotein complexes in the mushroom . The surfactant Triton X-100 played a critical role in the optimal method, and the targeting efficiency of the genomic editing reached 100% on a selective medium containing 5-FOA. This study is the first to use an RNP complex delivery to establish a CRISPR/Cas9 genome-editing system in . Moreover, compared with other methods, this method avoids the use of any foreign DNA, thus saving time and labor when it comes to plasmid construction.
基于体内表达的Cas9和向导RNA，在一些食用菌中建立了CRISPR/Cas9系统。与这些系统相比，体外组装的Cas9和sgRNA核糖核蛋白复合物（RNPs）具有更多优势，但仅有少数例子报道，且编辑效率相对较低。在本研究中，我们开发并优化了一种基于体外组装核糖核蛋白复合物的蘑菇CRISPR/Cas9基因组编辑方法。表面活性剂Triton X-100在该优化方法中起关键作用，在含有5-FOA的选择培养基上，基因组编辑的靶向效率达到了100%。本研究首次使用核糖核蛋白复合物递送在[具体物种未明确，原文此处缺失]中建立CRISPR/Cas9基因组编辑系统。此外，与其他方法相比，该方法避免使用任何外源DNA，因此在质粒构建方面节省了时间和人力。
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