In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline

Tran Thi Nhu Thao*, Fabien Labroussaa*, Nadine Ebert, Joerg Jores*, and Volker Thiel*
* contributed equally

In this book chapter a detailed protocol is provided for the assembly and engineering of infectious full-length coronavirus cDNAs using yeast-based transformation-associated recombination cloning. This novel assembly method not only results in stable coronavirus infectious full-length cDNAs cloned in the yeast Saccharomyces cerevisiae but also fosters and accelerates the manipulation of their genomes. The protocol is provided in readily-reproducible step-by step fashion, opening with an introductory overview, a list of the materials and reagents needed, followed by a detailed procedure that is supported with helpful notes offering tips and tricks of the trade as well as troubleshooting advice.

Printed August 2020 in Coronaviruses: Methods and Protocols.