Main description:

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.

Authoritative and cutting-edge, Yarrowia lipolytica: Methods and Protocols aims to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully.

Contents:

1. Genome Engineering of Yarrowia lipolytica with the piggyBac Transposon System

James M. Wagner, Claire M. Palmer, Maya V. Venkataraman, Lars H. Lauffer, Joshua M. Wiggers, Eden V. Williams, Xiunan Yi, and Hal S. Alper

2. Genome Editing in Y. lipolytica Using TALENs

Coraline Rigouin, Christian Croux, Gwendoline Dubois, Fayza Daboussi, and Florence Bordes

3. EasyCloneYALI: Toolbox for CRISPR-mediated Integrations and Deletions in Yarrowia lipolytica

Jonathan Dahlin, Carina Holkenbrink, and Irina Borodina

4. Simultaneous gene excision and integration by dual-guide CRISPR-Cas9

Michael Spagnuolo and Mark Blenner

5. Gene excision by dual-guide CRISPR-Cas9

Michael Spagnuolo and Mark Blenner

6. CRISPR Interference and Activation to Modulate Transcription in Yarrowia lipolytica

Joshua Misa and Cory Schwartz

7. Implementing CRISPR-Cas12a for Efficient Genome-editing in Yarrowia lipolytica

Zhiliang Yang and Peng Xu

8. Guide RNA Design for Genome-wide CRISPR Screens in Yarrowia lipolytica

Adithya Ramesh and Ian Wheeldon

9. Targeted Integration through Transformation of Hydroxyurea-Arrested Cells

Vasiliki Tsakraklides

10. Strain Construction for Intracellular Metabolic Pathway Localization in Y. lipolytica

Erin L. Bredeweg and Scott E. Baker

11. Title: Engineering Yarrowia lipolytica for production of fatty alcohols with YaliBrick Vectors

Wanqi Sun, Zhiliang Yang and Peng Xu

12. Methods to Activate and Elucidate Complex Endogenous Sugar Metabolism in Yarrowia lipolytica

Seunghyun Ryu and Cong T. Trinh

13. Pathway Engineering For Beta-Carotene and Carotenoid Biosynthesis in Y. lipolytica

Mauricio Pesantes-Munoz and Rodrigo Ledesma-Amaro

14. Droplet-based Microfluidic High-throughput Screening of Enzyme Mutant Libraries Secreted by Yarrowia lipolytica

Thomas Beneyton and Tristan Rossignol

15. Yarrowia lipolytica Bioprocess Development - From Flask to Bioreactor

Farshad Darvishi and Marzieh Moradi

16. Western Blotting of Membrane-Bound Proteins in Yarrowia lipolytica

Adithya Ramesh and Ian Wheeldon