Manipulating the Avian Egg: Applications for Embryo Transfer,Transgenics,and Cloning

In vitro production of germline chimeras and avian cloning may utilise the transfer of avian embryos from their original eggshell to a surrogate eggshell for culture during incubation. Such embryo transfer is valuable for avian cloning as the only alternative would be to transfer the cloned avian embryos into the infundibulum of recipient birds. Given the advances in paleogenomics, synthetic biology, and gene editing, a similar approach might be used to generate extinct species, i.e. de-extinction. One objective of the present research was to examine if ratite eggs could be manipulated via windowing and sham injection, similar to that which could allow for avian genome manipulation and subsequent development. The efficiency of interspecific avian embryo transfer using Chicken (Gallus gallus domesticus) donor eggs and Turkey (Meleagris gallopavo) recipient eggshells was also investigated. Egg windowing and embryo transfer techniques utilised in the present research were adapted from those found in the scientific literature. Presumed fertile eggs from Rhode Island Red (n = 40), Silkie (n = 2), and White Leghorn Chickens (n = 18), Turkey (n = 48), Emu (Dromaius novaehollandiae) (n = 79), and Ostrich (Struthio camelus) (n = 89) were used in this research. Of the 41 Chicken eggs used for transfers into recipient Turkey eggshells, only one (2.4%) produced a chick. Of 31 windowed Emu eggs, one embryo survived for 25 d but no chicks were produced. Of 36 windowed Ostrich eggs, one embryo survived and hatched. The efficiency of the windowing and embryo transfers to produce chicks was low and further refinements are needed. Importantly, the results herein establish that manipulating ratite embryos is possible.