Genome Engineering of Cryptococcus Neoformans using CRISPR-Cas System

 描述(由申请人提供)：新生隐球菌是一种机会性的人类病原体，在免疫缺陷的个人中会导致潜在的致命脑膜脑炎。新生假单胞菌的毒力是一个多方面的特征，它产生黑色素、多糖壳和分泌的有助于其毒力的蛋白质。最近引入的分子和基因技术使我们能够开始了解真菌毒力的分子基础，但由于缺乏先进的方法，精确的基因改变和高效的多重基因组编辑使系统地研究毒力机制仍然有限。我们建议开发一个CRISPR-Cas(集群规则间隔短回文重复序列-CRISPR相关)系统，用于对新生隐孢子虫进行精确的基因特异性修饰和高效的多重基因组编辑。这一创新的基因组方法的成功发展将为我们研究新生葡萄球菌的毒力机制提供一个革命性的工具。

 DESCRIPTION (provided by applicant): Cryptococcus neoformans is an opportunistic human pathogen that causes potentially fatal meningoencephalitis in individuals with immune deficiency. Virulence of C. neoformans is a multi-faceted trait with the production of the melanin pigment, a polysaccharide capsule, and secreted proteins contributing to its virulence. The recent introduction of molecular and genetic technologies have allowed us to begin to understand the molecular basis of fungal virulence, yet precision gene alterations and efficient multiplex genome editing that allows systematic examination of virulence mechanisms remain limited due to the lack of advanced approaches. We propose to develop a CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated) system for precise gene specific modification and efficient multiplex genome editing in C. neoformans. The successful development of this innovative genomic approach will provide a revolutionary tool in our studying of virulence mechanisms in C. neoformans.