Methods for Injecting Avian Eggs

Abstract

The present invention provides improved methods of injecting an avian egg containing an embryo, preferably an early embryo (e.g., a blastoderm). The methods of the invention may be used to deliver a substance to an egg, remove a sample from an egg, and/or to insert a detector device into an egg to collect information therefrom. In preferred embodiments, the invention is used to deliver a substance to the embryo in ovo. In other preferred embodiments, the invention is used to produce chimeric or transgenic avian embryos in ovo.

1 - 60 . (canceled)  
   
   
       61 . A method of producing a chimeric bird, comprising the steps of: 
 orienting an avian egg containing a blastoderm in a predetermined position;    introducing a small opening into a shell of the egg;    extending a delivery device through the small opening in the egg shell;    piercing an inner shell membrane with the delivery device, wherein the inner shell membrane is essentially intact prior to inserting the delivery device therethrough;    releasing an avian donor cell through the delivery device and depositing the donor cell into the blastoderm or in close proximity thereto under conditions sufficient to result in a chimeric embryo;    retracting the delivery device from the egg;    incubating the chimeric embryo to hatch to produce a chimeric bird.    
   
   
       62 . The method of  claim 61 , wherein the delivery device is extended through the small opening in the egg shell to a location selected from the group consisting of: 
 (a) the subgerminal cavity,    (b) between the area opaca and the vitelline membrane,    (c) between the area pellucida and the vitelline membrane,    (d) between the area opaca and the area pellucida,    (e) in the area pellucida,    (f) in the area opaca, and    (g) any combination of (a) to (f) above.    
   
   
       63 . A method of producing a transgenic bird, comprising the steps of: 
 orienting an avian egg containing a blastoderm in a predetermined position;    introducing a small opening into a shell of the egg;    extending a delivery device through the small opening in the egg shell; 
 piercing an inner shell membrane with the delivery device, wherein the inner shell membrane is essentially intact prior to inserting the delivery device therethrough;  
 releasing a substance comprising a nucleotide sequence through the delivery device and depositing the substance into the blastoderm or in close proximity thereto under conditions sufficient to result in a transgenic embryo;  
 retracting the delivery device from the egg;  
 incubating the transgenic embryo to hatch to produce a transgenic bird.  
   
   
   
       64 - 65 . (canceled)