Team:Valencia UPV
Covering food necessities is mandatory, but resources are not sustainably exploited. Global strategies to increase food productivity and quality need to be concealed with a local perspective, providing breeders with the necessary technology to improve varieties. The aim of HYPE-IT is to decrease current technological barriers for breeding local crops using precision genome engineering, easing the gene editing process using SynBio-inspired simplified CRISPR/Cas9 tools. HYPE-IT brings along a software tool that associates crop traits with specific gene targets and designs optimal gRNAs for those targets. HYPE-IT also incorporates a modular gene circuit that serves as an in vivo gRNA testing system, ensuring appropriate gRNA choice even when no precise sequence information of local varieties is available. We aim to develop a split-Cas9 system based on viral vectors to efficiently deliver the editing machinery into the plant, and to create an affordable Labcase with the necessary laboratory equipment for HYPE-IT.
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gRNA Testing System
The plant breeder could test the obtained guide RNA directly in his plant, but it would take a long time just to check if it works or not. To avoid this, we have designed a gRNA Testing System which does this in a fast and simple way. Using Nicotiana benthamiana, Agrobacterium infection system, a short sequence of the gene of his plant and a luciferase assay, the users will surely know if the guide RNA works on his own variety. This system works as a switch that will show luciferase if the gene has been knocked-out.
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Split Cas9
As a vehicle to genetically modify the plant, the Agrobacterium infection system is inefficient, long and difficult. Thus, we decide to use a viral system to enhance plant infection. Since the viral vectors are small, the Cas9 and CPF1 endonuclease - necessary for editing - need to be splitted in two parts. To reassemble them inside the plant cells, we will use intein proteins.
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Lab-Case
We want to make plant gene editing accessible for all potential users, even if they do not have an advanced laboratory. To achieve this goal, we have made a Lab-case, a hardware set up which contains all the laboratory equipment and the necessary reagents required to perform genome editing, with simple protocols. This guarantees availability for agricultural cooperatives, start-ups, garage laboratories... This will be the future of agriculture: making genome editing of plants accessible to everyone.
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