Mutagenesis & Physiology Overview
Injection of males with ENU introduces point mutations into their spermatogonial stem cells, such that on average new point mutations in any gene can be recovered in one out of 500 to 1000 progeny. We have developed high-throughput screening protocols for the analysis of all major organ systems by morphology, physiology, and pathology and are currently using these to identify dominant genome-wide mutations with clinically relevant phenotypes on the G1 ("first generation") progeny of the mutagenized G0 males.
Our current breeding schema involves mutagenized 129S1/SvImJ males mated to wild type C57BL/6J females. We are screening for defects in areas of current specialization by collaborating labs, including cardiovascular, hematopoietic, neurological and skeletal systems, in the same living mice.
Confirmed outliers are bred to assess the inheritance of the trait of interest - once heritability is confirmed then DNA from the progeny are sent to Dr. Lucy Osborne's lab at the University of Toronto (part of the TCAG) for chromosome localization. A minimum of 10 affected progeny is required. If a map position is found then the line is bred further for in-depth secondary screens by the collaborating lab and to narrow the map interval. Mutations are cloned through candidate gene analysis.
New collaborations and ideas for new screens are always welcomed.