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• W4377013963 abstract "RNA-templated/directed DNA repair is a biological mechanism that has been experimentally demonstrated in bacteria, yeast, and mammalian cells. Recent study has shown that small noncoding RNAs (DDRNAs) and/or newly RNAPII transcribed RNAs (dilncRNAs) are orchestrating the initial steps of double-strand break (DSB) repair. In this study, we demonstrate that also pre-mRNA could be used as direct or indirect substrate for DSB repair. Our test system is based on (1) a stably integrated mutant reporter gene that produces constitutively a nonspliceable pre-mRNA, (2) a transiently expressed sgRNA-guided dCas13b::ADAR fusion protein to specifically RNA edit the nonspliceable pre-mRNA, and (3) transiently expressed I-SceI to create a DSB situation to study the effect of spliceable pre-mRNA on DNA repair. Based on our data, the RNA-edited pre-mRNA was used in cis for the DSB repair process, thereby converting the genomically encoded mutant reporter gene into an active reporter gene. Overexpression and knockdown of several cellular proteins were performed to delineate their role in this novel RNA-mediated end joining pathway." @default.
• W4377013963 created "2023-05-19" @default.
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• W4377013963 date "2023-06-01" @default.
• W4377013963 modified "2023-10-16" @default.
• W4377013963 title "Model System to Analyze RNA-Mediated DNA Repair in Mammalian Cells" @default.
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• W4377013963 doi "https://doi.org/10.1089/crispr.2022.0105" @default.
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