The gene trap methodology is a powerful tool to characterize novel genes and analyze their importance in biological phenomena. It is based on the use of mouse embryonic stem cells and reporter vectors designed to randomly integrate into the genome, tagging an insertion site and generating a mutation. Theoretically, all the 100000 genes present in the mouse genome could be tagged and functionally inactivated at the same time. Here we describe the basic concepts and perspectives of this methodology and show some results obtained by the gene trap approach used to study molecular cascades in basic cell biology and in developmental processes. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.