Use of primordial germ cell based chicken transgene & knockout techniques to establish animal models for studies in development & infection

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The chicken is an excellent animal model for basic and applied research in animal sciences like developmental biology, physiology and immunology. As a consequence, the chicken genome was the first domestic animal genome to be sequenced. Based on this information, the identification of candidate genes in the different research fields and their functional characterisation is now possible. In the past, functional in vivo studies had been difficult to perform in chickens due to a lack of appropriate transgene technologies. Recent advances in long term culture methods of chicken primordial germ cells (PGC) have opened new ways to manipulate their genome and to generate transgenic and knock out birds. This project will first focus on the generation of PGCs from a highly inbred chicken strain called line 0. This line allows cell transfer and tissue transplantation studies without the risk of transplant rejection. Line 0 PGCs will be used for insertion of a DNA coding for a green fluorescent protein (eGFP). Green fluorescent transgenic PGCs will be injected into line 0 embryos to generate germ line transgenic birds. After mating for two generations the resulting offsprings will express eGFP in all cells. Fluorescent cells from these chickens will subsequently be used for cell transfer.The second aim of this project is to obtain gene knockout birds for additional research on chicken B cell biology. Using appropriate targeting vectors the light and the heavy chain of the B cell receptor will be knocked out in line 0 PGCs. Knock out birds derived from these PGCs will not be able to express a functional B cell receptor on precursor cells. As a consequence, these cells will not receive appropriate survival signals for further differentiation and will die prior to homing into the central organ for B cell development, the bursa of Fabricius. This work will lead to the first gene knock out bird published and to an excellent model for B cell research in birds.