An Efficient CRISPR/Cas9 Genome Editing System for a Ganoderma lucidum Cultivated Strain by Ribonucleoprotein Method
文献类型： 外文期刊
第一作者： Tan, Yi
作者： Tan, Yi;Feng, Na;Tang, Chuanhong;Zou, Gen;Zhang, Jingsong;Yu, Xianglin;Tian, Jialin;Zhang, Zhigang
作者机构：
关键词： Ganoderma lucidum; CRISPR/Cas9; genome editing; ribonucleoproteins (RNPs); the orotidine 5 & PRIME;-monophosphate decarboxylase gene (ura3)
期刊名称：JOURNAL OF FUNGI （ 影响因子：4.7； 五年影响因子：5.2 ）
ISSN：
年卷期： 2023 年 9 卷 12 期
页码：
收录情况： SCI
摘要： The CRISPR/Cas9 system has become a popular approach to genome editing. Compared with the plasmid-dependent CRISPR system, the ribonucleoprotein (RNP) complex formed by the in vitro assembly of Cas9 and single-guide RNA (sgRNA) has many advantages. However, only a few examples have been reported and the editing efficiency has been relatively low. In this study, we developed and optimized an RNP-mediated CRISPR/Cas9 genome editing system for the monokaryotic strain L1 from the Ganoderma lucidum cultivar 'Hunong No. 1'. On selective media containing 5-fluoroorotic acid (5-FOA), the targeting efficiency of the genomic editing reached 100%. The editing efficiency of the orotidine 5 '-monophosphate decarboxylase gene (ura3) was greater than 35 mutants/107 protoplasts, surpassing the previously reported G. lucidum CRISPR systems. Through insertion or substitution, 35 mutants introduced new sequences of 10-569 bp near the cleavage site of ura3 in the L1 genome, and the introduced sequences of 22 mutants (62.9%) were derived from the L1 genome itself. Among the 90 mutants, 85 mutants (94.4%) repaired DNA double-strand breaks (DSBs) through non-homologous end joining (NHEJ), and five mutants (5.6%) through microhomology-mediated end joining (MMEJ). This study revealed the repair characteristics of DSBs induced by RNA-programmed nuclease Cas9. Moreover, the G. lucidum genes cyp512a3 and cyp5359n1 have been edited using this system. This study is of significant importance for the targeted breeding and synthetic metabolic regulation of G. lucidum.
分类号：
- 相关文献
作者其他论文 更多>>
-
Self-assembled ferritin-based nanoparticles elicit a robust broad-spectrum protective immune response against SARS-CoV-2 variants
作者：Wang, Weiqi;Xia, Xianzhu;Wang, Weiqi;Meng, Xianyong;Wang, Shen;Feng, Na;Zhao, Yongkun;Wang, Tiecheng;Yan, Feihu;Xia, Xianzhu;Meng, Xianyong;Cui, Huan;Zhang, Cheng
关键词：SARS-CoV-2; Self -assembled ferritin; Nanoparticle vaccine; Cross -neutralizing antibody
-
Physicochemical properties of polysaccharides from Hericium erinaceus by steam explosion pretreatment and its effects on human gut microbiota
作者：Chen, Shuang;Zhang, Fangfang;Liu, Liping;Feng, Jie;Zhang, Jingsong;Yang, Yan;Wu, Di;Liu, Yanfang;Chen, Shuang;Zhang, Fangfang;Liu, Liping;Feng, Jie;Zhang, Jingsong;Yang, Yan;Wu, Di;Liu, Yanfang;Chen, Shuang;Zhang, Fangfang;Liu, Liping;Feng, Jie;Zhang, Jingsong;Yang, Yan;Wu, Di;Liu, Yanfang;Guo, Qingbin
关键词：Hericium erinaceus; Steam explosion; beta-glucan; In vitro digestion; Fecal fermentation; Gut microbiota
-
Current Advances in the Functional Genes of Edible and Medicinal Fungi: Research Techniques, Functional Analysis, and Prospects
作者：Li, Wenyun;Zou, Gen;Bao, Dapeng;Wu, Yingying;Li, Wenyun;Bao, Dapeng;Wu, Yingying
关键词：macrofungi; genetic transformation; gene editing; mating-type genes; active substances
-
A Bacterium-like Particle Vaccine Displaying Envelope Proteins of Canine Distemper Virus Can Induce Immune Responses in Mice and Dogs
作者：Liu, Lina;Liu, Lina;Li, Ranran;Wu, Jianzhao;Zhao, Yongkun;Yan, Feihu;Wang, Tiecheng;Gao, Yuwei;Zhao, Zongzheng;Feng, Na;Xia, Xianzhu;Wang, Jianzhong;Li, Ranran;Wu, Jianzhao;Feng, Na;Xia, Xianzhu
关键词：CDV; F protein; H protein; subunit vaccine; bacterium-like particles; pandas
-
A virus-like particle candidate vaccine based on CRISPR/Cas9 gene editing technology elicits broad-spectrum protection against SARS-CoV-2
作者：Wang, Weiqi;Xia, Xianzhu;Wang, Weiqi;Wang, Shen;Meng, Xianyong;Zhao, Yongkun;Li, Nan;Wang, Tiecheng;Feng, Na;Yan, Feihu;Xia, Xianzhu;Meng, Xianyong
关键词：CRISPR/Cas9; Canarypox-virus vector; Virus -like particle vaccine; Cross -neutralizing antibody; Cellular immunity; Toll -like receptor 4
-
Effects of Dietary Gracilaria lichenoides and Bacillus amyloliquefaciens on Growth Performance, Antioxidant Capacity, and Intestinal Health of Penaeus monodon
作者：Tian, Jialin;Yan, Hailiang;Tian, Jialin;Wang, Yun;Huang, Jianhua;Yan, Hailiang;Duan, Yafei;Wang, Jun;Zhou, Chuangpeng;Wang, Yun;Wang, Jun;Zhou, Chuangpeng;Huang, Zhong
关键词：Penaeus monodon; Gracilaria lichenoides; Bacillus amyloliquefaciens; growth; antioxidant capacity; intestine microbiota; lipid metabolism
-
Establishment of two serological methods for detecting IgG and neutralizing antibodies against Crimean-Congo hemorrhagic fever virus glycoprotein
作者：Wang, Qi;Xia, Xianzhu;Wang, Qi;Wang, Shen;Shi, Zhikang;Li, Zhengrong;Zhao, Yongkun;Feng, Na;Wang, Tiecheng;Yan, Feihu;Xia, Xianzhu;Li, Zhengrong
关键词：CCHFV; rGP; rVSV/CCHFV; ELISA and sVNT; specific antibodies and serological samples
