Jiménez García, Alberto

Ledesma Amaro, Rodrigo

Martínez Buey, Rubén

Muñoz‑Fernández, Gloria

Revuelta Doval, José Luis

English

[EN]The filamentous fungus Ashbya gossypii is currently used for the industrial production of vitamin B2. Furthermore, the ability of A. gossypii to grow using lowcost substrates together with the inexpensive downstream processing makes this fungus an attractive biotechnological chassis. Indeed, the production in A. gossypii of other high-added value compounds such as folic acid, nucleosides and biolipids has been described. Hence, the development of new methods to expand the molecular toolkit for A. gossypii genomic manipulation constitutes an important issue for the biotechnology of this fungus. In this work, we present a one-vector CRISPR/Cas9 system for genomic engineering of A. gossypii. We demonstrate the efficiency of the system as a marker-less approach for nucleotide deletions and substitutions both with visible and invisible phenotypes. Particularly, the system has been validated for three types of genomic editions: gene inactivation, the genomic erasure of loxP scars and the introduction of point mutations. We anticipate that the use of the CRISPR/Cas9 system for A. gossypii will largely contribute to facilitate the genomic manipulations of this industrial fungus in a marker-less mannerThis work was financed by grants from the Spanish Ministerio de Economía y Competitividad (BIO2014-56930-P and BIO2017-88435-R) and Junta de Castilla y León (SA016P17) to JLR and AJ

Gestion del Repositorio Documental de la Universidad de Salamanca

One-vector CRISPR/Cas9 genome engineering of the industrial fungus Ashbya gossypii

https://gredos.usal.es/bitstream/10366/150748/1/37.%20CRISPR_Microbial_Biotech.pdf

One-vector CRISPR/Cas9 genome engineering of the industrial fungus Ashbya gossypii

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Gestion del Repositorio Documental de la Universidad de Salamanca