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在Dictyostelium discoideum中基于CRISPR/Cas9的frataxin基因版本

Hernan G Gentili¹, María Florencia Pignataro¹, Justo Olmos¹

¹Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3), Departamento de Fisiología y Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes 2160, Ciudad Universitaria, C1428EGA Buenos Aires, Argentina.

The Biochemical journal

|September 18, 2023

概括

研究人员开发了一种缺乏frataxin (FXN) 的Dictyostelium discoideum菌株,以研究弗里德里希.

科学领域:

生物化学生化学
细胞生物学细胞生物学
遗传学是一个遗传学.

背景情况:

弗拉塔克辛 (FXN) 对于铁硫 (Fe-S) 集群生物发生至关重要.
缺少FXN会导致细胞功能障碍和疾病,包括弗里德里希的无氧症 (FA).
了解跨物种的FXN功能保护有助于疾病研究.

研究的目的:

创建和描述一个Dictyostelium discoideum FXN缺陷菌株.
为了研究人类和D. discoideum FXN.的功能性保存.
为了评估一个特定的人类FA FXN变体在D. discoideum.

主要方法:

在D. discoideum fxn locus. 的基因编辑.
隔离和表型分析一个fxn缺陷突变 (克隆8).
评估Fe-S集群依赖的酶活性,生长,氧化应激敏感性和多细胞发育.

主要成果:

突变D. discoideum fxn (克隆8) 显示Fe-S酶功能减弱,生长速度减慢,氧化应激敏感性增加.
突变的多细胞发育和细菌草生长受损.
人类FA FXN变种 (DdFXN-G122V) 部分挽救了发育缺陷,而不是完全挽救生长和酶活性.

结论:

缺少FXN的D. discoideum菌株作为研究FXN功能和FA的有价值模型.
该模型有助于探索多种FXN变体,并有助于治疗策略的药物查.
FXN的功能性保存突出了模型生物在人类疾病研究中的实用性.

关键词:

这种类型的体是Dictyostelium.弗拉塔克辛 (frataxin) 是一种弗里德里希·阿塔克西亚 (Friedreich Ataxia) 是一个铁硫集群组件组件组件

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