Fate Mapping

A great deal of interest has centered around the fate of individual cells within the developing embryo (29,30). In lower organisms, following the fate and genesis of cells is less complex than in a mammal such as the mouse embryo, where marking a single cell by injection or transplantation and following its descendants during the course of development is technically demanding, requiring expensive equipment and expertise.

ES cell-mediated transgenic technologies utilizing the Cre/loxP system may be able to facilitate fate mapping the embryo greatly. Here a lineage-restricted promoter is used to drive the Cre recombinase. A second transgene containing a reporter gene flanked by the loxP sites is also required. The second transgene should contain a "stop" sequence between the ubiquitous promoter and the marker gene to keep the gene silent. Double transgenic animals will neither express Cre nor the marker gene until the specific developmental stage permitting Cre expression and subsequent recombination, resulting in marker gene activation. This will result in activation of the recombinase, resulting in the excision of the "stop" sequence, and expression of the marker in all the progenitors cells, regardless of the later expression status of the Cre-driving promoter. If conditions are optimized, then the Cre can be used for noncomplete excision and, thus, result in excision in a very limited number of cells or even in single cell. Another possible improvement would be the utilization of an inducible Cre recombinase, allowing the precise regulation of excision frequency and timing, thereby making it relatively straightforward to follow the fate of individual cells and their descendants based on the expression pattern of the marker.

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