TY - JOUR
T1 - DIVERSIFY: A Fungal Multispecies Gene Expression Platform
AU - Jarczynska, Zofia D.
AU - Rendsvig, Jakob K. H.
AU - Pagels, Nichlas
AU - Viana, Veronica R.
AU - Nødvig, Christina S.
AU - Kirchner, Ferdinand H.
AU - Strucko, Tomas
AU - Nielsen, Michael L.
AU - Mortensen, Uffe H.
PY - 2021
Y1 - 2021
N2 - Recent sequencing of numerous fungal species revealed large repertoires of putative biotechnologically relevant genes and secondary metabolite gene clusters. However, often the commercial potential of these species is impeded by difficulties to predict host physiological and metabolic compatibility with a given product, and lack of adequate genetic tools. Consequently, most heterologous production is performed in standard hosts where genetic tools and experience are in place. However, these species may not be suitable for all products. To increase chances of successful heterologous production, we have created a flexible platform, DIVERSIFY, for multispecies heterologous gene expression. This reduces the workload to construction of a single gene expression cassette, used to transform all DIVERSIFY strains in order to identify the optimal cell factory host. As proof of principle of the DIVERSIFY concept, we present the first version of our platform, DIVERSIFY 1.0, which we have successfully used for the production of three proteins and a metabolite in four different Aspergilli species, and for the identification of the best producer for each of the products. Moreover, we show that DIVERSIFY 1.0 is compatible with marker-free gene targeting induced by the CRISPR nucleases Cas9 and MAD7.
AB - Recent sequencing of numerous fungal species revealed large repertoires of putative biotechnologically relevant genes and secondary metabolite gene clusters. However, often the commercial potential of these species is impeded by difficulties to predict host physiological and metabolic compatibility with a given product, and lack of adequate genetic tools. Consequently, most heterologous production is performed in standard hosts where genetic tools and experience are in place. However, these species may not be suitable for all products. To increase chances of successful heterologous production, we have created a flexible platform, DIVERSIFY, for multispecies heterologous gene expression. This reduces the workload to construction of a single gene expression cassette, used to transform all DIVERSIFY strains in order to identify the optimal cell factory host. As proof of principle of the DIVERSIFY concept, we present the first version of our platform, DIVERSIFY 1.0, which we have successfully used for the production of three proteins and a metabolite in four different Aspergilli species, and for the identification of the best producer for each of the products. Moreover, we show that DIVERSIFY 1.0 is compatible with marker-free gene targeting induced by the CRISPR nucleases Cas9 and MAD7.
KW - Aspergillus
KW - Cell factories
KW - Gene expression platform
KW - CRISPR
KW - Cas9
KW - MAD7
U2 - 10.1021/acssynbio.0c00587
DO - 10.1021/acssynbio.0c00587
M3 - Journal article
C2 - 33651591
VL - 10
SP - 579
EP - 588
JO - A C S Synthetic Biology
JF - A C S Synthetic Biology
SN - 2161-5063
IS - 3
ER -