@article{Bonturi:2022:10.1016/j.mec.2022.e00200,
author = {Bonturi, N and Pinheiro, MJ and de, Oliveira PM and Rusadze, E and Eichinger, T and Liudit, G and De, Biaggi JS and Brauer, A and Remm, M and Miranda, EA and Ledesma-Amaro, R and Lahtvee, P-J},
doi = {10.1016/j.mec.2022.e00200},
journal = {Metabolic Engineering Communications},
pages = {1--10},
title = {Development of a dedicated Golden Gate Assembly Platform (RtGGA) for Rhodotorula toruloides.},
url = {http://dx.doi.org/10.1016/j.mec.2022.e00200},
volume = {15},
year = {2022}
}

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TY  - JOUR
AB  - Rhodotorula toruloides is a potential chassis for microbial cell factories as this yeast can metabolise different substrates into a diverse range of natural products, but the lack of efficient synthetic biology tools hinders its applicability. In this study, the modular, versatile and efficient Golden Gate DNA assembly system (RtGGA) was adapted to the first basidiomycete, an oleaginous yeast R. toruloides. R. toruloides CCT 0783 was sequenced, and used for the GGA design. The DNA fragments were assembled with predesigned 4-nt overhangs and a library of standardized parts was created containing promoters, genes, terminators, insertional regions, and resistance genes. The library was combined to create cassettes for the characterization of promoters strength and to overexpress the carotenoid production pathway. A variety of reagents, plasmids, and strategies were used and the RtGGA proved to be robust. The RtGGA was used to build three versions of the carotenoid overexpression cassette by using different promoter combinations. The cassettes were transformed into R. toruloides and the three new strains were characterized. Total carotenoid concentration increased by 41%. The dedicated GGA platform fills a gap in the advanced genome engineering toolkit for R. toruloides, enabling the efficient design of complex metabolic pathways.
AU  - Bonturi,N
AU  - Pinheiro,MJ
AU  - de,Oliveira PM
AU  - Rusadze,E
AU  - Eichinger,T
AU  - Liudit,G
AU  - De,Biaggi JS
AU  - Brauer,A
AU  - Remm,M
AU  - Miranda,EA
AU  - Ledesma-Amaro,R
AU  - Lahtvee,P-J
DO  - 10.1016/j.mec.2022.e00200
EP  - 10
PY  - 2022///
SN  - 2214-0301
SP  - 1
TI  - Development of a dedicated Golden Gate Assembly Platform (RtGGA) for Rhodotorula toruloides.
T2  - Metabolic Engineering Communications
UR  - http://dx.doi.org/10.1016/j.mec.2022.e00200
UR  - https://www.ncbi.nlm.nih.gov/pubmed/35662893
UR  - https://www.sciencedirect.com/science/article/pii/S2214030122000098?via%3Dihub
UR  - http://hdl.handle.net/10044/1/98595
VL  - 15
ER  - 

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