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Study of the in vivo function of sry, a key element in mammalian sex determination

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The testis determining gene Sry is a key element in the mammalian sex-determination pathway. There is some in vitro evidence that the mouse SRY protein acts as a transcriptional activator. To test this hypothesis in vivo, we will create a repressor allele of SRY in transgenic mice. An active repressor can be generated by linking the transcription repressor domain of the Drosophila gen Engrailed to the binding domain of SRY. If this causes sex reversal in the transgenic mice, then SRY probably acts as a transcriptional activator. The reverse experiment, adding a strong activator domai to SRY, should prove this.
The cellular distribution of SRY has never been determined in vivo, because good antibodies are not yet available. We propose to create an epitope tagged SRY protein in transgenic mice. Using antibodies against the epitope, we will be able to detect the protein in vivo. The existence of an epitope tagged SRY protein will also aid in the search for downstream target genes.

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Medical Research Council
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