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Profile nucleases and Repurpose Off-Targets to Expand Gene Editing

Project description

Using off-target gene editing to advantage

Gene editing relies on making a targeted double-strand break in a sequence of interest. ‘Programmable nucleases’ found in microbes do just this. They can degrade the DNA of viruses that infect microbes (phages), while leaving microbial DNA intact. There are thousands of programmable nucleases in microbial genomes, but too little is known about their actions. The lack of ‘guide-target parity’ (cutting only fully matched target genome sites) is a significant concern. The ERC-funded PROTEGE project aims to turn this into an advantage. The team will use high-throughput biochemistry to profile nucleases for the specificity of guide-target parity, create a toolset to assess this, and test whether purposefully ‘misprogramming’ nucleases can direct specific repair outcomes.

Objective

Programmable nucleases reside at the nexus of advanced gene editing and microbial defense. These nucleases degrade phage DNA but avoid genomic DNA to provide the microbe with adaptive immunity. The standard-bearer, CRISPR-Cas9, relies on a guide RNA (gRNA) – its program – to precisely cut a complimentary genetic sequence – its target – for precision gene editing. These programmable nucleases deliver cure for genetic diseases like anemia, blindness, and cancer; quick disease model development; and diagnostics for viruses like SARS-CoV-2.
Hunts across microbial genomes have exposed thousands of programmable nucleases. Each could be another valuable tool for genome engineers, but we know just too little about them to use them safely. Even Cas9 does not have guide-target parity: It cuts partially matched ‘off-target’ genome sites, which can lead to dangerous side effects. Though we work hard to avoid off-targets, partial matching may have a bright side. What if we leverage it, using it to make gene editing more predictable?
Using high-throughput biochemistry, we will Profile nucleases and Repurpose Off-Targets to Expand Gene Editing. The PROTÉGÉ research program will profile guide-target parity – specificity – across promising, newly discovered programmable nucleases. It will deliver a toolset for assessing this critical safety feature rapidly and broadly. Finally, we will test the hypothesis that purposefully misprogramming nucleases can direct specific repair outcomes. We intended to benefit Europe and the world community with safer and more diverse gene editing tools for achieving its Horizon Europe health, technology and environmental goals.

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HORIZON-ERC - HORIZON ERC Grants

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Call for proposal

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(opens in new window) ERC-2022-STG

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Host institution

VILNIAUS UNIVERSITETAS
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 1 141 779,00
Address
UNIVERSITETO G. 3
01513 Vilnius
Lithuania

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Region
Lietuva Sostinės regionas Vilniaus apskritis
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 1 141 779,00

Beneficiaries (1)

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