EasyCloneYALI: Toolbox for CRISPR-Mediated Integrations and Deletions in Yarrowia lipolytica

In order to unlock the full potential of Yarrowia lipolytica, as model organism and production host, simple and reliable tools for genome engineering are essential. In this chapter, the practical details of working with the EasyCloneYALI Toolbox are described.Highlights of the EasyCloneYALI Toolbox are high genome editing efficiencies, multiplexed Cas9-mediated knockouts, targeted genomic integrations into characterized intergenic loci, as well as streamlined and convenient cloning for both marker-based and marker-free integrative expression vectors.