Bibliographic Details
| Title: |
HEK293T Cells with TFAM Disruption by CRISPR-Cas9 as a Model for Mitochondrial Regulation. |
| Authors: |
de Oliveira, Vanessa Cristina1 (AUTHOR) kroballo@vt.vcom.edu, Santos Roballo, Kelly Cristine1,2,3 (AUTHOR) clesio.gmm@usp.br, Mariano Junior, Clésio Gomes1 (AUTHOR) sarahingrid@usp.br, Santos, Sarah Ingrid Pinto1 (AUTHOR) fabianabressan@usp.br, Bressan, Fabiana Fernandes1 (AUTHOR) ceambrosio@usp.br, Chiaratti, Marcos Roberto4 (AUTHOR) chiarattimr@gmail.com, Tucker, Elena J.5,6 (AUTHOR) elena.tucker@mcri.edu.au, Davis, Erica E.7,8,9 (AUTHOR) eridavis@luriechildrens.org, Concordet, Jean-Paul10 (AUTHOR) jean-paul.concordet@mnhn.fr, Ambrósio, Carlos Eduardo1 (AUTHOR) |
| Source: |
Life (2075-1729). Jan2022, Vol. 12 Issue 1, p22-N.PAG. 1p. |
| Subject Terms: |
*CRISPRS, *MITOCHONDRIAL DNA, *GENOME editing, *MITOCHONDRIA, *STAINS & staining (Microscopy), *TRANSCRIPTION factors |
| Abstract: |
The 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. [ABSTRACT FROM AUTHOR] |
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