VISCREEN · Engineering viral vectors for high-throughput CRISPR genetic screens in plants
Horizon Europe — Marie Skłodowska-Curie Actions
- Duration
- 2023-04-01 → 2025-03-31
- EU contribution
- €175,920
- Participants
- 2
- Scheme
- HORIZON-TMA-MSCA-PF-EF
Lines connect the coordinator with its partners.
Results in brief
Engineering viral vectors for high-throughput CRISPR genetic screens in plants
CRISPR-Cas genome editing offers new opportunities to carry out genetic screens and understand fundamental concepts in plant genetics. The simultaneous delivery of thousands of guide RNAs (gRNAs) into cells allows to rapidly create an ordered mutant collection overcoming functional redundancy in plant genomes (mainly due to polyploidization and genome duplications). While standard CRISPR screens rely on plasmid pools delivered via conventional transformation techniques, the delivery of CRISPR-Cas components using plant viral vectors offers a promising alternative for a rapid and efficient modification without genetic transformation. Combining virus-induced genome editing (VIGE) with CRISPR represents a novel approach to generate large mutant populations in non-transformable cultivars. The VISCREEN project aims to develop and expand the current toolbox of viral vectors so that VIGE can be implemented as a powerful, easy-to-use tool for plant genome engineering. I will approach two specific objectives: (1) develop a high-throughput genome editing system using Barley stripe mosaic virus (BSMV) for wheat and maize; and (2) engineer a set of viral vectors for gene editing in soybean.
Data: CORDIS, © European Union
Project objective
Tools based on CRISPR-Cas systems hold great potential both for fundamental plant genetics and precision crop breeding. Unlike traditional mutagenic approaches, CRISPR-Cas systems allow for the simultaneous delivery of thousands of unique guide RNAs, enabling the generation of collections of higher-order mutants for rapid functional screens and the discovery of genetic interactions. However, CRISPR technology suffers from a ‘transformation bottleneck’; relatively few crop genotypes can be transformed and often only a handful of mutant lines can be produced. In virus-induced genome editing (VIGE), plant viruses are engineered to deliver gRNAs into cells. Such a capability is highly attractive for CRISPR knock-out (KO) screens as it could simplify the delivery and generate large populations of mutant lines in a broader range of crop genotypes. However, the application of VIGE for CRISPR KO screens is hampered by the lack of a proper viral-based multiplex gRNA delivery vector. My host lab has pioneered the development of CRISPR KO screens in plants, and in VISCREEN I will combine this knowledge with my expertise on VIGE to scale up CRISPR-Cas-mediated manipulation of plant genomes. The main objectives of my project are: (i) to develop Barley stripe mosaic virus (BSMV) for high-throughput CRISPR screens in wheat and maize; and (ii) to engineer and validate a set of viral vectors for VIGE in soybean. The viral vector-based, high-throughput editing systems developed in VISCREEN will expand CRISPR screens in a wider variety of crops and allow to test fundamental biological questions that are limited by functional redundancy. Overall, this innovative approach will make me a well-trained academic researcher with the capabilities to develop a professional career within the plant genome editing field.
Original text from CORDIS.
Participants
Links
- View on CORDIS
- DOI: 10.3030/101110621
- https://ec.europa.eu/research/participants/documents/downloadPublic?documentIds=080166e505002391&appId=PPGMS
Data: CORDIS, © European Union