Yarrowia lipolytica Methods and Protocols

This volume provides a collection of protocols for the most common experimental methods used for engineering Yarrowia lipolytica. Chapters detail the basic theories underlying the methods described in each chapter. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Genome Engineering of Yarrowia lipolytica with the piggyBac Transposon System.- Genome Editing in Y. lipolytica Using TALENs.- EasyCloneYALI: Toolbox for CRISPR-mediated Integrations and Deletions in Yarrowia lipolytica.- Simultaneous gene excision and integration by dual-guide CRISPR-Cas9.- Gene excision by dual-guide CRISPR-Cas9.- CRISPR Interference and Activation to Modulate Transcription in Yarrowia lipolytica..- Implementing CRISPR-Cas12a for Efficient Genome-editing in Yarrowia lipolytica.- Guide RNA Design for Genome-wide CRISPR Screens in Yarrowia lipolytica.- Targeted Integration through Transformation of Hydroxyurea-Arrested Cells.- Strain Construction for Intracellular Metabolic Pathway Localization in Y. lipolytica.- Engineering Yarrowia lipolytica for production of fatty alcohols with YaliBrick Vectors.- Methods to Activate and Elucidate ComplexEndogenous Sugar Metabolism in Yarrowia lipolytica.- Pathway Engineering For Beta-Carotene and Carotenoid Biosynthesis in Y. lipolytica.- Droplet-based Microfluidic High-throughput Screening of Enzyme Mutant Libraries Secreted by Yarrowia lipolytica.- Yarrowia lipolytica Bioprocess Development – From Flask to Bioreactor.- Western Blotting of Membrane-Bound Proteins in Yarrowia lipolytica.