Experiments to Study DNA Double-Strand Break Repair by RNA in Human Cells
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Zhang, Yiqi
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Abstract
In this study, research was conducted on DNA double-strand breaks (DSBs) and the role of RNA in their repair mechanisms. While efforts to prevent DSB formation face challenges, understanding repair pathways like homologous recombination (HR), non-homologous end joining (NHEJ), and microhomology-mediated end joining (MMEJ) is crucial. Recent studies suggest RNA may serve as an alternative repair template alongside DNA and mediate DSB repair by other repair mechanisms. This study utilized CRISPR-Cas9 cleavage systems to induce specific DSBs on an exon on the DsRed gene in human cells and evaluated the direct involvement of RNA in DSB repair on the exonic region. In vitro cleavage assays and in vivo experiments with HEK293T cells were conducted, analyzing repair frequencies and mechanisms. Results indicate that RNA's impact on repair differs based on DSB location and RNA transcript levels.
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2024-04-29
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