de Oliveira, Vanessa CristinaSantos Roballo, Kelly CristineMariano Junior, Clesio GomesSantos, Sarah Ingrid PintoBressan, Fabiana FernandesChiaratti, Marcos RobertoTucker, Elena J.Davis, Erica E.Concordet, Jean-PaulAmbrosio, Carlos Eduardo2022-08-242022-08-242022-0122http://hdl.handle.net/10919/111618The mitochondrial transcription factor A (TFAM) is considered a key factor in mitochondrial DNA (mtDNA) copy number. Given that the regulation of active copies of mtDNA is still not fully understood, we investigated the effects of CRISPR-Cas9 gene editing of TFAM in human embryonic kidney (HEK) 293T cells on mtDNA copy number. The aim of this study was to generate a new in vitro model by CRISPR-Cas9 system by editing the TFAM locus in HEK293T cells. Among the resulting single-cell clones, seven had high mutation rates (67-96%) and showed a decrease in mtDNA copy number compared to control. Cell staining with Mitotracker Red showed a reduction in fluorescence in the edited cells compared to the non-edited cells. Our findings suggest that the mtDNA copy number is directly related to TFAM control and its disruption results in interference with mitochondrial stability and maintenance.application/pdfenCreative Commons Attribution 4.0 InternationalCRISPR-Cas9gene editingHEK293T cellsmitochondrial DNATFAMArticle - Refereedhttps://doi.org/10.3390/life12010022121350544162075-1729