GNRECC · Gene network rewiring by an engineered CRIPSR-Cas12a variant
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2020-09-01 → 2022-08-31
- EU contribution
- €171,473
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Gene network rewiring by an engineered CRIPSR-Cas12a variant
A major obstacle to the understanding of cellular physiology in both health and disease arise from the ability of genes to execute cellular programs. Therefore, how these elements are interconnected is still an open question which limits our comprehension of complex cell behaviors. To elucidate and control the gene network underlying cellular functions, this project aims to engineer Cas12a, a member of CRISPR/Cas protein family used for cell transcriptional reprogramming. Although highly potent in single-gene control, the efficacy of Cas12a decreases when directed to multiple genes. Therefore, more potent Cas12a version are needed to facilitate rapid and large-scale engineering of cellular programs. By designing novel Cas12a enzyme able to be directed towards multiple genomic loci simultaneously, this project will simplify the generation of novel genetic pathways useful for the biotechnological and industrial sector including biopharmaceuticals production and cell-based therapies. Therefore, we aim at combining statistical inference techniques with both protein and genome engineering to address current challenges that limit the explosion of gene network engineering field. This project aims to generate a potent Cas12a useful for multiplexed orthogonal gene control and employ this optimized enzyme for the validation of an inferred molecular network. To address this issue, the following work packages (WPs) will be executed: rational design of novel Cas12a variants (WP1), validation of novel Cas12a variants (WP2), gene network rewiring by a novel Cas12a variant (WP3).
Data: CORDIS, © European Union
Project objective
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.
Original text from CORDIS.
Participants
- POLITECNICO DI TORINO · TorinoCoordinatorItaly
Links
- View on CORDIS
- DOI: 10.3030/894574
- https://www.researchers.polito.it/en/success_stories/marie_sklodowska_curie_individual_fellowships/rapid_easy_and_efficient_gene_network_reprogramming
Data: CORDIS, © European Union