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Development of a new CRISPR Cas3 based tool for large genomic deletions

The advent of the revolutionary genome editing technique CRISPR-Cas9 has enabled targeted gene mutation, repression, and activation, facilitating impactful biological findings. However, Cas9 as an unbiased DNA deletion tool is lim... ver más

30/06/2022

EMBL

265K€

Presupuesto del proyecto: 265K€

Líder del proyecto

EUROPEAN MOLECULAR BIOLOGY LABORATORY No se ha especificado una descripción o un objeto social para esta compañía.

Fecha límite participación Sin fecha límite de participación.

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Financiación concedida El organismo H2020 notifico la concesión del proyecto el día 2022-06-30

MSCA-IF-2018: Individual Fellowships

I+D

Cerrada hace 6 años

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3 Participantes

EMBL

264.67K€ | Lider

THE REGENTS OF THE UNIVERSIT...

LOS ANGELES UCLA SANTA BARBA...

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Duración del proyecto: 38 meses Fecha Inicio: 2019-04-08
Fecha Fin: 2022-06-30

Líder del proyecto

EUROPEAN MOLECULAR BIOLOGY LABORATORY No se ha especificado una descripción o un objeto social para esta compañía.

Presupuesto del proyecto 265K€

Fecha límite de participación Sin fecha límite de participación.

Descripción del proyecto The advent of the revolutionary genome editing technique CRISPR-Cas9 has enabled targeted gene mutation, repression, and activation, facilitating impactful biological findings. However, Cas9 as an unbiased DNA deletion tool is limited in its ability to interrogate large regions of DNA of unknown function, because it predominantly generates very small (<20 bp) insertions and deletions at its target site. The capacity to rapidly and efficiently generate large genomic deletions does not currently exist and would be an extremely useful tool for research, allowing for rapid strain engineering of bacterial cells for synthetic biological and metabolic engineering purposes. Additionally, this technology would allow for the interrogation of large segments of non-coding DNA in human cells, much of which has unknown function, but whose variants are often associated with human disease. In this proposal, I aim to develop a Type I-C CRISPR-Cas system employing the Cas3 enzyme (completely distinct from Cas9), which naturally possess coupled nuclease and helicase activity, for high-throughput gene-editing purposes in various prokaryotic, as well as human cells. My preliminary results have shown that this is a credible approach, as I have been able to generate individually, as well as in combination, multiple deletions in bacterial organisms exceeding 60 kb in size. A focal point of the proposal is to adapt this system for use in human cells, which would provide a novel basic research tool with unprecedented capabilities and also could be utilized in human health-related applications. The proposal aims to address this later possibility by utilizing the developed system to treat human cell lines infected with difficult-to-treat pathogenic viruses.

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