Afg3l1^tm1.1Arte

An FRT-flanked neo cassette with a 5' loxP site was inserted upstream of exon 2. An additional loxP site was inserted downstream of exon 3. Cre mediated recombination removed the FRT-flanked neo cassette and exons 2 and 3. RT-PCR analysis of liver tissue indicated residual mRNAs after splicing from exon 1 to either exon 4 or exon 5. While exon 1-4 splicing gives rise to an out-of-frame transcript, splicing from exon 1 to 5 leads to an in-frame transcript potentially encoding a shorter protein that is devoid of large part of the mitochondrial targeting sequence. However, immunoblotting of liver mitochondria revealed no mature protein in homozygous mutant mice. Blue-native PAGE showed lack of assembled protein in high-molecular weight m-AAA complexes, confirming that this is a knock-out allele. (J:237410)