Defining the test space of complementation assays

To systematically evaluate yeast/human cross-species complementation in the context of human disease, the test space was defined as follows: first, all yeast/human orthologous pairs were extracted from the InParanoid database (http://inparanoid.sbc.su.se/cgi-bin/index.cgi) (Ostlund et al. 2010). Pairs not meeting all of the following four criteria were eliminated: (1) human genes must have a disease-associated variant identified in either OMIM or HGMD; (2) human genes must have had an open reading frame (ORF­) clone available in the Human ORFeome v8.1 library (Yang et al. 2011); (3) yeast genes must be essential (Winzeler et al. 1999; Giaever et al. 2002); and (4) yeast genes must have an available temperature-sensitive (ts) mutant (Ben-Aroya et al. 2008; Li et al. 2011). In addition to published ts strains, 36 additional strains were obtained through mutagenesis and screening (G Tan, BJ Andrews, and C Boone, unpubl.).