Genome Engineering of Cryptococcus Neoformans using CRISPR-Cas System

 描述（由申请方提供）：新型隐球菌是一种机会性人类病原体，可在免疫缺陷个体中引起潜在致死性脑膜脑炎。C.新型原虫是一种多方面的性状，产生黑色素、多糖囊和有助于其毒力的分泌蛋白。最近引入的分子和遗传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，精确的基因改变和有效的多重基因组编辑，允许系统检查毒力机制仍然有限。我们建议开发一种CRISPR-Cas（规则间隔短回文重复序列-CRISPR相关）系统，用于精确的基因特异性修饰和高效的C.新人类这种创新的基因组方法的成功开发将为我们研究C.新人类

 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.