Partial phenotypic rescue of the Sesb1 mitochondrial ANT1 disease model in Drosophila

Study of gene expression patterns of Drosophila melanogaster Sesb1 mutants compared to wild type Inbreeding under stressful conditions inevitably results in the selection of compensatory alleles of many genes. In order to avoid such issues, and thus determine the global effects on gene expression of the Sesb1 mutation in a truly unselected and 'wild-type' background, we outbred Sesb1 over more than 10 generations by back-crossing to each of two commonly used wild-type strains, Canton S and Oregon R. Subsequent to this backcrossing, Sesb1 was maintained in each background using a balancer chromosome. These stocks were then used to generate a Sesb1 mutant F1 generation, by crossing virgin Canton S Sesb1. homozygous mutant females with Oregon R Sesb1 males. For comparison, we generated otherwise isogenic wild-type F1 progeny by crossing virgin Canton S wild-type females with Oregon R wild-type males. Progeny was collected on the day of eclosion and aged for one day after which the RNA was isolated with Trizol. The quality of the RNA was analyzed with a spectrophotometer.