A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families.

A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families.
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白色念珠菌CRISPR系统允许基因和基因家族的基因工程。

DOI:
10.1126/sciadv.1500248
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发表时间:
2015
期刊:
影响因子:
13.6
通讯作者:
Fink GR
Fink GR
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Vyas VK;Barrasa MI;Fink GR

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CRISPR 简化了人类真菌病原体白色念珠菌的基因工程。白色念珠菌是一种致病性酵母菌,可引起粘膜和系统感染,死亡率很高。缺乏简便的分子遗传学一直是发病机制分析的主要障碍。减数分裂的缺乏加上质粒的缺乏使得基因工程变得繁琐,特别是对于基本功能和基因家族而言。我们描述了一种白色念珠菌 CRISPR 系统,该系统克服了该生物体基因工程的许多障碍。 CRISPR 诱导的突变可以高频率定向到目标基因,即使不进行选择,也可以轻松分离纯合基因敲除。此外,该系统允许创建在多个基因、基因家族和编码基本功能的基因中具有突变的菌株。该 CRISPR 系统对于未确定倍性的新鲜临床分离株也有效。我们的方法改变了操纵念珠菌基因组的能力,并为了解这种病原体的生物学提供了一个新的窗口。
CRISPR simplifies genetic engineering of the human fungal pathogen Candida albicans. Candida albicans is a pathogenic yeast that causes mucosal and systematic infections with high mortality. The absence of facile molecular genetics has been a major impediment to analysis of pathogenesis. The lack of meiosis coupled with the absence of plasmids makes genetic engineering cumbersome, especially for essential functions and gene families. We describe a C. albicans CRISPR system that overcomes many of the obstacles to genetic engineering in this organism. The high frequency with which CRISPR-induced mutations can be directed to target genes enables easy isolation of homozygous gene knockouts, even without selection. Moreover, the system permits the creation of strains with mutations in multiple genes, gene families, and genes that encode essential functions. This CRISPR system is also effective in a fresh clinical isolate of undetermined ploidy. Our method transforms the ability to manipulate the genome of Candida and provides a new window into the biology of this pathogen.
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