245249 20230208174206.0 G:(EU-Grant)894574 894574 CORDIS G:(EU-Call)H2020-MSCA-IF-2019 H2020-MSCA-IF-2019 CORDIS corda__h2020::894574 originalID G:(EU-Grant)894574 Gene network rewiring by an engineered CRIPSR-Cas12a variant 2020-09-01 - 2022-08-31 H2020-MSCA-IF-2019 2020-09-01 2022-08-31 GNRECC d 2020-09-01 - 2022-08-31 I:(DE-588b)5098525-5 European Union CORDIS In the last 10 years, an increasing number of studies employed -omics technologies to describe the molecular changes underpinning cellular functions. However, a large part of these findings is awaiting an experimental validation. This is due to the lack of efficient molecular tools able to control both multiple and distinct genetic interactions. Recently, I described a new CRISPR/12-based genome engineering tool that, for the first time, it provides the constitutive, conditional, inducible, orthogonal and multiplexed engineering of dozens of endogenous genes, simultaneously. In this research proposal, I aim to increase the efficiency of this platform in the context of multiplexed genome engineering applications, such as gene network rewiring. To this aim, I will use techniques inspired by the statistical physics of complex disordered systems, to design a more potent version of my CRISPR/Cas12-based genome engineering tool and, I will use this novel platform to rewire signaling pathways involved in cellular proliferation. This research proposal is structured into the following tasks: rational design of novel Cas12a variants, validation of novel Cas12a variants, gene network rewiring by a novel Cas12a variant. By coupling statistical physics to both protein and genome engineering this project will pave the way for efficient and large-scale engineering of gene networks. oai:juser.fz-juelich.de:901689 authority:GRANT authority G CORDIS AUTHORITY oai:juser.fz-juelich.de:901689