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Expanding the genome editing toolbox by rational reprogramming of tyrosine recombinase DNA specificities

Project description

Establishing tyrosine recombinases as advanced tools for genome editing

For safety and effectiveness, gene therapies often need to integrate a large therapeutic DNA cargo at precise genomic locations. In this case, DNA editors such as CRISPR technology are limited by nuclease-based mechanisms and cannot adequately handle large DNA insertions. Meanwhile, tyrosine recombinases are capable of rearranging large DNA segments without nuclease-based mechanisms. However, there is no current solution for rational reprogramming of their DNA specificities for binding non-native DNA targets. The ERC-funded EditYR project aims to create a rationally programmable tyrosine recombinase platform that enables efficient integration of large DNA cargo into precise genomic locations. It will extend recombinase engineering capabilities by leveraging recent advances in large-scale DNA synthesis and sequencing along with bioinformatics and protein design.

Objective

Gene therapies aim to cure genetic diseases by modifying the DNA blueprint. An ideal gene therapy should be durable, safe, and efficient, which often requires integration of large therapeutic DNA cargo at precise genomic locations. Advanced DNA editors with programmable specificities, such as the CRISPR technology, are limited by the nuclease-based mechanism and poor efficiency of large DNA insertions. Increased efficiency, specificity and precision of large edits are required to address a broad therapeutic space.
Tyrosine recombinases (YRs) efficiently rearrange large DNA segments without the pitfalls of nuclease-based editors, but with an important caveat – rational reprogramming of their specificities for binding non-native DNA targets is an unsolved problem. EditYR ambitiously aims to develop a rationally programmable tyrosine recombinase platform for efficient integration of large DNA cargo into precise genomic locations. EditYR will expand recombinase engineering capabilities by exploiting the recent advances in large scale DNA synthesis and sequencing coupled with bioinformatics and innovative protein design. Objective 1 will engineer tyrosine recombinase DNA specificities and extract a comprehensive DNA recognition code. Objective 2 will modify their oligomerization properties to enable binding non-palindromic targets, allowing targeting any desired nucleotide sequence. Objective 3 will investigate requirements for irreversible recombination to maximize integration efficiencies and unlock the full therapeutic potential of these enzymes. Finally, Objective 4 will validate engineered enzymes by identification of targetable sequences in therapeutically relevant genomic regions and integration of therapeutic DNA cargo in various human cell types for different disease cases.EditYR will establish tyrosine recombinases as advanced tools for genome editing, creating a DNA editing technology that will elevate the capabilities of gene therapies to unprecedented levels

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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-2024-STG

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Host institution

KEMIJSKI INSTITUT
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.

€ 1 599 500,00
Address
HAJDRIHOVA 19
1000 Ljubljana
Slovenia

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Region
Slovenija Zahodna Slovenija Osrednjeslovenska
Activity type
Research Organisations
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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.

€ 1 599 500,00

Beneficiaries (1)

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