Swine in Biomedical Research Conference 2005, Chicago, IL. Abstract GC7.
Comparative genome information for humans and pigs is well established and provides the basis for comparative phenotypic maps between rodents, humans and pigs. Significant progress has been achieved in the analysis of phenotypes (functional genomics) using gene-driven mutagenesis approaches for the functional analysis of the mouse genome. We are using DNA-sequence together with gene targeting technologies to resolve complex traits (diseases) in pigs. This approach is based on the hypothesis that recombineering and gene-trapping in porcine fetal fibroblasts coupled with animal cloning through nuclear transfer permits development of biomedical models and provide insights into gene function. These models can support the use of comparative phenomics between mouse, rat, dog, man, and the pig. We have developed a recombineering platform to introduce targeted changes in porcine fibroblast and other somatic cells. We have been able to target deletions and insertions as well as introduce selection markers into specific genes. In addition, we have reduced the targeted BAC from 180 kb to 20 kb by gap repair rescue methods which increased transfection efficiency in the fetal fibroblasts (FF). Through gene-trapping of ES cells, a collection of random mutant lines can be developed as the Bay Genomics project (NIH-sponsored) has done. They focus on a mouse ES gene-trap model for deciphering new gene function associated with cardiopulmonary development and related diseases. Thus, we are using gene-trapping technology in porcine fetal fibroblasts FF to develop cell lines for defining phenotypes. Both recombineering and gene-trapping approaches will provide core sources for addressing biomedical models of complex traits.