基于CRISPR的工具用于致病性Sporothrix物种的有针对性的基因操纵
Remi Hatinguais1, Ian Leaves1, Gordon D Brown1
1Medical Research Council Centre for Medical Mycology, University of Exeter , Exeter, United Kingdom.
Microbiology spectrum
|September 14, 2023
概括
研究人员开发了一个新的CRISPR/Cas9基因编辑工具,用于Sporothrix真菌,使得有针对性的基因删除. 这有助于我们更好地了解运动三病的毒性因素,并有助于开发新的治疗方法.
科学领域:
- 菌类学和传染病的研究
- 分子生物学和遗传学
背景情况:
- 斯波托里克斯巴西利尼斯 (Sporothrix brasiliensis) 致使斯波托里科斯 (Sporotrichosis),这是一个新兴的动物性疾病,在拉丁美洲迅速蔓延.
- 有限的分子工具阻碍了对Sporothrix毒性因子和疾病机制的研究.
- 了解真菌病原体对于开发有效的诊断和治疗至关重要.
研究的目的:
- 建立一种强大的基因操纵方法,用于Sporothrix物种.
- 为了研究毒性因素,使得有针对性的基因删除成为可能.
- 克服Sporothrix研究的逆遗传学方面的局限性.
主要方法:
- 开发了一种用于异质基因表达的PEG介导的原质细胞转化协议.
- 使用了CRISPR/Cas9基因编辑与转换协议相结合.
- 删除了假定的DHN-黑色素合成酶基因 (pks1) 和KU80同类基因,以增强同类重组.
主要成果:
- 在S. brasiliensis,S. schenckii和S. chilensis中成功证明了异质基因表达.
- 启用了pks1基因的向删除,这是一个拟议的毒性因子.
- 删除KU80增强了同源定向修复,提高了CRISPR/Cas9.9的向基因删除效率.
结论:
- 开发的基于CRISPR/Cas9的转换协议是针对Sporothrix物种的向基因操纵的一个有效工具.
- 这一进步解决了对Sporothrix的可靠基因工程工具的稀缺问题.
- 促进研究毒性决定因素及其在子三病原发生的作用.
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