New approaches to develop in vivo model systems that detect somatic and germ line mutations (report II)

Abstract

We plan to develop new assay systems that use GFP for mutant detection. The principle of the approach is to make the cells fluorescent when mutation occurred at specified gene loci in the genome. One example is to introduce cells co-expression of two vectors, one produces GFP constitutively and the other produces repressor protein to silence the expression of GFP gene. In this system, any kinds of forward mutation in the repressor gene allow GFP gene transcription from null state, which should make the mutant cells easy to be recognized by green fluorescence. Another example is to make partial duplications of endogenous gene, in which the distal segment of the duplicate has a GFP gene tag. In this system, reversion from the duplicate makes the mutant cells fluorescent. Toward the generation of mouse model systems, we have been evaluating the systems in cultured cells and measuring spontaneous and radiation-induced mutants by flow cytometry.