A unified multi-kingdom Golden Gate cloning platform

Chiasson D, Giménez-Oya V, Bircheneder M, Bachmaier S, Studtrucker T, Ryan J, Sollweck K, Leonhardt H, Boshart M, Dietrich P, Parniske M (2019)

Publication Type: Journal article

Publication year: 2019

Journal

Scientific Reports Nature Publishing Group: Open Access Journals - Option B

Book Volume: 9

Article Number: 10131

Journal Issue: 1

DOI: 10.1038/s41598-019-46171-2

Abstract

Assembling composite DNA modules from custom DNA parts has become routine due to recent technological breakthroughs such as Golden Gate modular cloning. Using Golden Gate, one can efficiently assemble custom transcription units and piece units together to generate higher-order assemblies. Although Golden Gate cloning systems have been developed to assemble DNA plasmids required for experimental work in model species, they are not typically applicable to organisms from other kingdoms. Consequently, a typical molecular biology laboratory working across kingdoms must use multiple cloning strategies to assemble DNA constructs for experimental assays. To simplify the DNA assembly process, we developed a multi-kingdom (MK) Golden Gate assembly platform for experimental work in species from the kingdoms Fungi, Eubacteria, Protista, Plantae, and Animalia. Plasmid backbone and part overhangs are consistent across the platform, saving both time and resources in the laboratory. We demonstrate the functionality of the system by performing a variety of experiments across kingdoms including genome editing, fluorescence microscopy, and protein interaction assays. The versatile MK system therefore streamlines the assembly of modular DNA constructs for biological assays across a range of model organisms.

Authors with CRIS profile

Tanja Studtrucker Professur für Zellbiologie der Pflanzen Petra Dietrich Professur für Zellbiologie der Pflanzen

Involved external institutions

Ludwig-Maximilians-Universität (LMU) DE Germany (DE)

How to cite

APA:

Chiasson, D., Giménez-Oya, V., Bircheneder, M., Bachmaier, S., Studtrucker, T., Ryan, J.,... Parniske, M. (2019). A unified multi-kingdom Golden Gate cloning platform. Scientific Reports, 9(1). https://dx.doi.org/10.1038/s41598-019-46171-2

MLA:

Chiasson, David, et al. "A unified multi-kingdom Golden Gate cloning platform." Scientific Reports 9.1 (2019).

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