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The project Crispy is dedicated to the development of an innovative toolbox to promote genome editing in the yeast Y. lipolytica, so as to establish it as an industrial platform organism. 


Y. lipolytica is a GRAS organism used in a wide variety of industrial applications to secrete or accumulate valuable metabolites (citric acid, natural or functionalized lipids …). Although some basic genetic tools are available, development of dedicated strains as industrial biofactories has been hampered by the paucity of genome engineering tools. The classical approach is based on homologous recombination, which allows gene insertion at a specific locus and targeted gene deletion. Although this methodology has been successfully employed in the past for rewriting lipid metabolism (Oleovision & ProBio3) or creating new metabolic pathways, it is nevertheless time consuming when multiple gene deletions have to be performed. Engineering of essential enzyme activities and evolution of multiple enzyme systems are impossible using this current approach. This project will provide an innovative toolbox using a targeting nuclease-based strategy for genome editing and will strengthen Yarrowia as an industrial platform organism. This new approach opens the way towards synthetic biology strategies. 


The project  is based on the complementarity of 3 partners. The groups of the LISBP and Micalis have been collaborating for more than 10 years on different projects combining Enzyme and Metabolic engineering of this yeast for the synthesis of high-value products (bio-kerosene, ricinoleic acid, bioplastic). Their knowledge of the yeast physiology, genetics, genomics and tools for enzyme engineering is crucial for the project. The development of groundbreaking technology for genome editing in this yeast will also benefit from the recent arrival in the LISBP of a leader in these technologies (Dr Fayza Daboussi). 

This project was funded in 2016 in the context of the precompetivie call for projects of TWB.

Florence Bordes: TBI "Catalyse et ingénierie moléculaire enzymatiques, Cimes"
Fayza Daboussi : TBI « Biologie de synthèse chez les microalgues, BioSyntMic " 
Cécile Neuveglise : MICALIS, " Diversité génomique et fonctionnelle des levures, DivY
Delphine Lestrade TWB 



Ce projet est co-financé par de la Région Occitanie / Pyrénées-Méditerranée et par le FEDER/FSE (programme 2014-2020).