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The careful comparison of the phenotypic variations generated by different alleles at a given locus, including of course, those alleles with a deleterious effect, is often an important source of information for the understanding of gene functions. In fact, every time it is possible to match a specific alteration observed at the genomic level with a particular pathology, it is possible to establish a relationship between a gene and its function. When considered from this point of view, the production of new mutations by experimental mutagenesis appears as an alternative to the strategy of in vitro gene invalidation by homologous recombination in embryonic stem (ES) cells, with the advantage that experimental mutagenesis does not require any previous knowledge of the gene structure at the molecular level. Homologous recombination in ES cells is a 'gene driven' approach, in which mutant alleles are produced for those genes that we already know. Experimental mutagenesis, on the contrary, is a 'phenotype driven' approach, in which unknown genes are identified based on phenotypic changes. Also, while homologous recombination in ES cells requires a rather sophisticated technology, mutagenesis is simple to achieve but relies greatly on the efficiency of the mutagenic treatment as well as on the use of an accurate protocol for phenotyping. In this review, we will address a few comments about the different techniques that can be used for the induction of point mutations in the mouse germ line with special emphasis on chemical mutagenesis. We will also discuss the limitations of experimental mutagenesis and the necessity to look for alternative ways for the discovery of new genes and gene functions in the mouse.

Reverse transcriptase catalyses the conversion of RNA into DNA. This operation seems to have largely contributed to the evolution of complex genomes. More than 10% of a mammalian genome is composed of sequences with reverse transcribed origin, most of which consists of repeated sequences (SINEs, LINEs). In spite of their simplicity, these sequences can play a key role in evolution by favoring illegitimate recombination. In addition to this abundant material, retrotransposed sequences include retrotransposons, retroviruses and genes depleted from intervening sequences, known as pseudogenes. Some of these sequences can be functional or involved in the regulation of neighbouring genes. These hallmarks of reverse transcription activity indicate that it has largely contributed to the fluidity of modern genomes.