Candida albicans is the leading cause of fungal infections; yet, complex genetic interaction analysis remains cumbersome in this diploid pathogen. Here, we developed a CRISPR-Cas9-based 'gene drive array' platform to facilitate efficient genetic analysis in C. albicans. In our system, a modified DNA donor molecule acts as a selfish genetic element, replaces the targeted site and propagates to replace additional wild-type loci. Using mating-competent C. albicans haploids, each carrying a different gene drive disabling a gene of interest, we are able to create diploid strains that are homozygous double-deletion mutants. We generate double-gene deletion libraries to demonstrate this technology, targeting antifungal efflux and biofilm adhesion factors. We screen these libraries to identify virulence regulators and determine how genetic networks shift under diverse conditions. This platform transforms our ability to perform genetic interaction analysis in C. albicans and is readily extended to other fungal pathogens.

中文翻译：

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基于 CRISPR-Cas9 的基因驱动平台，用于白色念珠菌遗传相互作用分析。


白色念珠菌是真菌感染的主要原因；然而，对于这种二倍体病原体，复杂的遗传相互作用分析仍然很麻烦。在这里，我们开发了一个基于 CRISPR-Cas9 的“基因驱动阵列”平台，以促进白色念珠菌的高效遗传分析。在我们的系统中，经过修饰的 DNA 供体分子充当自私的遗传元件，替换目标位点并传播以替换其他野生型基因座。使用具有交配能力的白色念珠菌单倍体，每个单倍体携带不同的基因驱动，使感兴趣的基因失效，我们能够创建纯合双缺失突变体的二倍体菌株。我们生成双基因删除文库来演示这项技术，针对抗真菌外排和生物膜粘附因子。我们筛选这些文库以识别毒力调节因子并确定遗传网络在不同条件下如何变化。该平台改变了我们在白色念珠菌中进行遗传相互作用分析的能力，并且很容易扩展到其他真菌病原体。