Genetic manipulation of spermatogonia

Abstract

The invention features a method of delivering DNA to a spermatogonium by infusing DNA in situ into a testicle of a non-human animal and administering a condition or substance to the testicle to increase uptake of DNA by the spermatogonium

What is claimed is:  
     
         1 . A method of delivering a DNA to a spermatogonium, comprising infusing in situ DNA into a testicle of a non-human animal and administering a condition or substance to said testicle to increase uptake of said DNA by said spermatogonium.  
     
     
         2 . The method of  claim 1 , wherein said condition is passage of an electrical current through the testicle.  
     
     
         3 . The method of  claim 2 , wherein said electrical current is applied to said testicle using a defibrillator.  
     
     
         4 . The method of  claim 2 , wherein said electrical current is applied to said testicle using an elect roej aculator.  
     
     
         5 . The method of  claim 1 , wherein said substance is a lipid or a phospholipid.  
     
     
         6 . The method of  claim 1 , wherein said DNA is infused in a volume of at least 0.1 ml per testicle.  
     
     
         7 . The method of  claim 1 , wherein the epididymis is surgically exposed at the head and the DNA is delivered to the testes via a retrograde flush through the rete testes into the seminiferous tubules.  
     
     
         8 . The method of  claim 1 , wherein said DNA is introduced into said spermatogonium by viral infection.  
     
     
         9 . The method of  claim 1 , wherein said non-human animal is selected from the group consisting of a sheep, goat, pig, cow, chicken, rabbit, rat, mouse, and guinea pig.  
     
     
         10 . The method of  claim 9 , wherein said non-human animal is a pig.  
     
     
         11 . The method of  claim 1 , wherein said animal is prepubetal.  
     
     
         12 . The method of  claim 1 , wherein said DNA comprises a sequence encoding a selectable marker.  
     
     
         13 . The method of  claim 12 , wherein said selectable marker is selected from the group consisting of antibiotic resistance gene, a cell surface antigen, or thymidine kinase.  
     
     
         14 . The method of  claim 1 , wherein DNA is administered to said testicle before the time at which sperm production is detected.  
     
     
         15 . The method of  claim 9 , wherein the age of said pig is at least 30 days.  
     
     
         16 . The method of  claim 15 , wherein the age of said pig is not greater than 100 days.  
     
     
         17 . The method of  claim 1 , wherein said DNA is naked.  
     
     
         18 . A method of making a non-human transgenic animal comprising infusing in situ DNA into a testicle of a prepubetal non-human animal, harvesting sperm cells from said animal, contacting an ovum with said sperm cells under conditions suitable for fertilization to produce said non-human transgenic animal.  
     
     
         19 . The method of  claim 18 , wherein said prepubetal non-human animal is a pig.  
     
     
         20 . The method of  claim 18 , wherein said pig is at least 30 days but not greater than 100 days of age.