Gene targeting of primordial germ cells using TALEN technology to generate the first knockout transgenic chickens

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The chicken is used to study how embryos form and is also a major agricultural species. Being able to selectively target genes in chickens will help to understand and improve disease resistance in birds and poultry production. This will also give us new ways to study how chicken embryos form. Sex stem cells can be used to precisely modify the chicken genome. Gene targeting using TALE nucleases (TALENs) promises to be an efficient method to modify sex stem cells to make knockout chickens. We propose to develop this novel technology to efficiently target genes in sex stem cells using TALEN-directed homologous recombination. In this proposal, we will target two genes needed for germ cells to form. Targeted sex stem cells will be injected into host embryos and bred to produce targeted knockout chickens. The knockout chicken produced will be useful to understand how germ cells form and also as a recipient for sex stem cells from frozen avian biobanks for domesticated poultry and potentially for endangered avian species. These advances will support research in basic biology of vertebrate development with applications in understanding disease resistance and fertility in poultry.