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| Generation of Mitochondrial Transcription Factor a Knockdown Transgenic Mice |
| QIN Yao, ZHAO Hong-yan, ZHANG Wen-hang, WANG Dong-mei |
| Affiliated Hospital of Zunyi Medical Colleague, Zunyi 563003, China |
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Abstract Objective: To produce mitochondria deficiency transgenic mice, providing a useful animal model for the investigation of mitochondria related diseases. Methods: Transgenic mice were produced by injection of mitochondrial transcription factor A (TFAM) RNAi lentivirus into zygotes. Frozen sections of tissues wre prepared to observe GFP expression. Variations of TFAM mRNA and protein were separately detected by quantitative PCR (qPCR) and western blot. Changes of mtDNA copy number were measured by qPCR. Results: We constructed lentiviral vectors expressing shRNA targeting TFAM, which markedly repressed the transcription and translation of TFAM in transduced MEF cells. TFAM knockdown mice were successfully produced by infection of the purified virus into the perivitelline space of single-cell embryo. Green fluorescence was observed in heart, liver, spleen, lung and kidney of transgenic mice. Using qPCR and western blot analysis, we found that the transcription of TFAM in the tissues of transgenic mice was significantly decreased. Moreover, reduced expression of TFAM in myocardium led to a decline of mitochondrial DNA (mtDNA) copy number, as well as an impaired respiratory chain function. Conclusion: A TFAM knockdown transgenic mice line was successfully constructed.
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Received: 22 April 2014
Published: 25 July 2014
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