Descripción del proyecto
Una nueva tecnología para editar grandes segmentos de ADN
La reciente llegada de la tecnología CRISPR-Cas9 ha revolucionado la edición del genoma dirigida al generar pequeñas inserciones y deleciones en el sitio de destino. El proyecto GenDels, financiado con fondos europeos, trabajará en una tecnología alternativa basada en CRISPR que emplea la enzima Cas3, que combina la actividad de la helicasa y la nucleasa. El objetivo es lograr una edición de genes de alto rendimiento de segmentos grandes de ADN en células humanas para aplicaciones relacionadas con la salud, o descifrar el papel de las variantes de ADN no codificantes en la enfermedad. Además, esta tecnología podría aprovecharse para editar células bacterianas con fines sintéticos biológicos y metabólicos.
Objetivo
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.
Ámbito científico (EuroSciVoc)
CORDIS clasifica los proyectos con EuroSciVoc, una taxonomía plurilingüe de ámbitos científicos, mediante un proceso semiautomático basado en técnicas de procesamiento del lenguaje natural.
CORDIS clasifica los proyectos con EuroSciVoc, una taxonomía plurilingüe de ámbitos científicos, mediante un proceso semiautomático basado en técnicas de procesamiento del lenguaje natural.
- ciencias naturalesciencias biológicasmicrobiologíavirología
- ciencias naturalesciencias biológicasgenéticaADN
- ciencias naturalesciencias biológicasgenéticamutación
- ciencias naturalesciencias biológicasgenéticagenoma
- ciencias naturalesciencias biológicasbioquímicabiomoléculasproteínasenzima
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Régimen de financiación
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinador
69117 Heidelberg
Alemania