Fig. 1. Creation and verification of doxycycline-inducible mtDNA-depleter mice.

a Alignment of amino acid sequences of polymerase domain of POLG1 protein from Homo sapiens to Neurospora crassa shows that aspartic acid in the POLG1 at 1135 position is evolutionarily conserved. b Schematic of the development of inducible D1135A-POLG1 (mtDNA-depleter) transgenic mouse model. D1135A-POLG1-expressing mouse (Mouse I) was created by microinjection of the pTRE-Tight-BI-AcGFP1-D1135-POLG1 construct into the one-cell stage egg from C57BL/6 mouse. The D1135A-POLG1-positive founder male mouse (Mouse I) was bred with the CAG-rtTA3 female mouse (Mouse II, Jackson Laboratories, stock # 016532) to get the D1135A-POLG1 transgenic animal (Mouse III). c Pups genotyping reveals the presence of D1135A-POLG1, rtTA, and GFP. d Whole-body imaging also confirms expression of GFP in only mtDNA-depleter mice. e RT-PCR analyses confirm dox-dependent expression of D1135A-POLG1 in only mtDNA-depleter mice