Deleting ku80 improves the efficiency of targeted gene editing in Neospora caninum

IF 1.4 4区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular and biochemical parasitology Pub Date : 2022-09-01 DOI:10.1016/j.molbiopara.2022.111508

Kaijian Wu , Xingju Song , Yayun Wu , Xu Yang , Jing Liu , Qun Liu

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引用次数: 0

Abstract

CRISPR/Cas9 technology has been widely used for gene editing in organisms. Gene deletion of the ku80/ku70 complex can improve the efficiency of gene replacement in Arabidopsis thaliana, Cryptococcus neoformans, and Toxoplasma gondii, which remained elusive in Neospora caninum. Here, we knock out the ku80 gene in Nc1 strain by using CRISPR/Cas9, detect the growth rate and virulence of NcΔku80. Then we compare the efficiency of gene replacements between NcΔku80 and Nc1 strains by transfected with the same HA-tagged plasmids, and the percentage of HA-tagged parasites was investigated by IFA. The results showed that gene targeting efficiency was increased in the NcΔku80 strain via double crossover at several genetic loci, but its growth rate and virulence were unaffected. In conclusion, the NcΔku80 strain can be used as an effective strain for rapid gene editing of N. caninum.

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删除ku80提高了犬新孢子虫靶向基因编辑的效率

CRISPR/Cas9技术已广泛应用于生物体内的基因编辑。ku80/ku70复合体的基因缺失可以提高拟南芥、新隐球菌和刚地弓形虫的基因替换效率，而这在犬新孢子虫中尚不明确。我们利用CRISPR/Cas9敲除Nc1株的ku80基因，检测NcΔku80的生长速度和毒力。然后我们比较了NcΔku80和Nc1菌株用相同ha标记的质粒转染后的基因置换效率，并用IFA法研究了ha标记的寄生虫的百分比。结果表明，通过多个基因位点的双杂交，NcΔku80菌株的基因靶向效率提高，但其生长速度和毒力未受影响。结果表明，NcΔku80菌株可作为一种高效的犬乳杆菌快速基因编辑菌株。

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来源期刊

Molecular and biochemical parasitology 医学-寄生虫学

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

63 days

期刊介绍： The journal provides a medium for rapid publication of investigations of the molecular biology and biochemistry of parasitic protozoa and helminths and their interactions with both the definitive and intermediate host. The main subject areas covered are: • the structure, biosynthesis, degradation, properties and function of DNA, RNA, proteins, lipids, carbohydrates and small molecular-weight substances • intermediary metabolism and bioenergetics • drug target characterization and the mode of action of antiparasitic drugs • molecular and biochemical aspects of membrane structure and function • host-parasite relationships that focus on the parasite, particularly as related to specific parasite molecules. • analysis of genes and genome structure, function and expression • analysis of variation in parasite populations relevant to genetic exchange, pathogenesis, drug and vaccine target characterization, and drug resistance. • parasite protein trafficking, organelle biogenesis, and cellular structure especially with reference to the roles of specific molecules • parasite programmed cell death, development, and cell division at the molecular level.