大型水蚤
生物
异型生物质的
百草枯
突变体
戒毒(替代医学)
细胞色素P450
基因
单加氧酶
遗传学
毒性
生物化学
化学
新陈代谢
酶
有机化学
替代医学
病理
医学
作者
Pijar Religia,Nhan Duc Nguyen,Quang Dang Nong,Tomoaki Matsuura,Yasuhiko Kato,Hajime Watanabe
摘要
Abstract The freshwater crustacean Daphnia magna has traditionally been a model for ecotoxicological studies owing to its sensitivity to many xenobiotics. Because it is used in many toxicity assessments, its detoxification mechanism for xenobiotics is important and requires further study. However, studies related to detoxification genes are limited to transcriptomic profiling, and there are no D. magna mutants for use in the understanding of xenobiotic metabolism in vivo. We report the generation of a D. magna CYP360A8 mutant—the gene is a cytochrome P450 (CYP) clan 3 gene. Based on RNA sequencing of adult D. magna , we found that CYP360A8 has the highest expression level among all CYP genes. At ovarian maturation, its expression level is up‐regulated 6‐fold compared to the juvenile stages and is maintained thereafter. Using the CRISPR/CRISPR‐associated 9 (Cas9) system, we disrupted CYP360A8 by coinjecting CYP360A8 ‐targeting guide RNA and Cas9 proteins into D. magna eggs and established one monoallelic CYP360A8 mutant line. This CYP360A8 mutant had a higher sensitivity to the herbicide paraquat compared to the wild type. We confirmed the up‐regulation of CYP360A8 by paraquat. The results demonstrate the role of CYP360A8 in paraquat detoxification. The present study establishes a CYP mutant of D. magna , and this strategy can be a basic platform to document a range of CYP gene–xenobiotic relationships in this species. Environ Toxicol Chem 2021;40:1279–1288. © 2020 SETAC
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