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| Dissertation / PhD Thesis | DKFZ-2024-02608 |
2024
University of Heidelberg
Heidelberg
Please use a persistent id in citations: https://katalog.ub.uni-heidelberg.de/titel/69220146
Abstract: Programmable RNA-guided technologies have sparked a revolution in both basic and translational research in the natural sciences. The discovery and development of the RNA-guided CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas9 system has made genome editing more accessible and broadly applicable. Its applications include, among others, characterization of gene functions, identification of disease-associated genes, and development of novel gene editing therapies. The sequence-specificity of the CRISPR-Cas system has also proven to be an invaluable tool in molecular diagnostics, where it allows the detection of specific bacterial or viral sequences during an infection. CRISPR-based diagnostic (CRISPR-Dx) technologies have been promptly applied to detect SARS-CoV-2 during the COVID-19 pandemic. In Chapter 1, I describe my contribution to this field by reporting the step-by-step optimization of a sensitive, rapid, and adaptable COVID-19 diagnostic test called ADESSO (Accurate Detection of Evolving SARS-CoV-2 through SHERLOCK Optimization) for the detection of SARS-CoV-2 and its variants.Other programmable RNA-guided technologies exploit the activity of the Adenosine Deaminase Acting on RNA (ADAR) enzymes. ADARs can be recruited by a guide RNA (gRNA) to a desired RNA sequence and induce specific adenosine to inosine (A-to-I) nucleotide changes. In Chapter 2, I describe an application of targeted ADAR-mediated RNA editing to regulate the immunogenicity of an epitope. The work described here paves the way for its application as a strategy to generate cancer neoepitopes in tumors to increase their immunogenicity and favor responsiveness to immunotherapy.
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