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Identification of engrailed target genes


Research objectives and content:
Engrailed-1/2 is a homeoprotein that plays an important role in mid-hindbrain development. My goal is to identify genes under Engrailed-1/2 transcriptional control, to study their expression and analyze their function.
To this end W. Wurst and A. Prochiantz have designed a gene trap strategy in which the LacZ gene integrated in the genome of ES cells is used as a reporter. The screen is based on the capability of the
Engrailed-2-homeodomain (En-2HD) internalized by ES cells to antagonize the activity of endogenous Engrailed-1/2 and modify the transcription of Engrailed-1/2-controlled genes. This approach has already allowed to characterize Dystonin as a true Engrailed-1/2 target. I will characterize three Engrailed-1/2 responding ES cell clones by identifying the corresponding genes, to study their expression and functional role. In a complementary approach, based on the internalization of En-2HD by midbrain-hindbrain neurons, I will use differential RNA display (DRD) to identify putative targets of Engrailed-1/2 involved at later stages of neurogenesis. In practice, I have already performed the DRD which have led to the cloning of several differential displayed PCR fragments that need to be fully characterized to initiate functional studies.
Training content (objective, benefit and expected impact):
During my postdoctoral stay I will benefit from the expertise of my host laboratory to complete my theoretical and practical training in developmental neurobiology. The collaboration with W. Wurst's laboratory in Munich (GSF) will allow me to generate homozygous mouse mutants in the best conditions and will give me access to the already existing En-l-- and En-2-- mice. The contact with the latter outstanding laboratory in my home-country will facilitate the planning of my future scientific career.


46,Rue D' Ulm 46 Ecole Normale Superieure
75230 Paris

Participants (1)

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