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

Tran, Thi Nhu Thao; Labroussaa, Fabien; Ebert, Nadine; Jores, Jörg; Thiel, Volker (2020). In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline. Methods in molecular biology, 2203, pp. 167-184. Springer 10.1007/978-1-0716-0900-2_13

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The Escherichia coli and vaccinia virus-based reverse genetics systems have been widely applied for the manipulation and engineering of coronavirus genomes. These systems, however, present several limitations and are sometimes difficult to establish in a timely manner for (re-)emerging viruses. In this chapter, we present a new universal reverse genetics platform for the assembly and engineering of infectious full-length 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. Such a platform is widely applicable for the scientific community, as it requires no specific equipment and can be performed in a standard laboratory setting. The protocol described can be easily adapted to virtually all known or emerging coronaviruses, such as Middle East respiratory syndrome coronavirus (MERS-CoV).

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology
05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Tran, Thi Nhu Thao; Labroussaa, Fabien; Ebert, Nadine; Jores, Jörg and Thiel, Volker Earl

Subjects:

600 Technology > 630 Agriculture
500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1940-6029

Publisher:

Springer

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

08 Apr 2021 09:00

Last Modified:

08 Apr 2021 09:00

Publisher DOI:

10.1007/978-1-0716-0900-2_13

PubMed ID:

32833212

Uncontrolled Keywords:

Coronavirus Full-length cDNA clone Homologous recombination RNA virus Reverse genetics Saccharomyces cerevisiae Synthetic genomics Transformation-associated recombination (TAR) cloning

BORIS DOI:

10.7892/boris.152072

URI:

https://boris.unibe.ch/id/eprint/152072

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