在二形真菌病原体的基因删除中使用优化后期CRISPR/Cas9系统进行向的基因删除
1Department of Microbiology, Ohio State University , Columbus, Ohio, USA.
mSphere
|June 30, 2023
概括
这项研究介绍了一种优化的CRISPR/Cas系统,用于在Histoplasma (一种真菌病原体) 中有效编辑基因. 这一突破加速了逆转基因研究,使得基因缺失的标记物减少.
科学领域:
- 分子生物学分子生物学
- 菌类学 菌类学是指菌类学.
- 遗传学 遗传学是一种遗传学.
背景情况:
- 组织等离子体是一种重要的真菌病原体,具有有限的逆遗传工具.
- 有效的基因失活对于理解质体毒性机制至关重要.
研究的目的:
- 开发和优化CRISPR/Cas系统,以有效地编辑血清蛋白中的基因.
- 为了促进基因删除的产生,并加速对这种病原体的逆遗传学研究.
主要方法:
- 使用单个表达指导RNA (gRNA) 和Streptococcus pyogenes Cas9.的情节性向量开发了一个优化的CRISPR/Cas系统.
- 双双接的gRNAs来自强大的Pol (II) 促进体,并由 ribozymes 进行处理.
- 聚合分离物的基于PCR的选使得可以隔离缺乏标记物的删除突变.
主要成果:
- 在CRISPR/Cas系统有效地产生了基因删除在各种质体物种.
- 成功隔离了没有标记的删除突变.
- 该系统对具有可选择和不可选择表型的多个基因进行了验证.
结论:
- 优化的CRISPR/Cas系统显著提高了对质体进行逆转基因的能力.
- 这种工具有望加速对Histoplasma spp.中的基因功能和毒性机制的研究.
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