Multitransgenic pigs comprising ten genetic modifications for xenotransplantation

A method of making a transgenic pig comprising at least six transgenes comprising the step of:

• (i) transfecting a porcine cell with a single polycistronic vector comprising SEQ ID NO:
• 12;
• (ii) producing a multitransgenic porcine cell comprising at least six transgenes by incorporating and expressing the polycistronic vector at a single genomic locus;
• (iii) generating a multitransgenic porcine zygote by injecting the nucleus of the multitransgenic porcine cell into a reconstructed somatic cell nuclear transfer (SCNT);
• and
• (iv) permitting the multitransgenic porcine zygote to mature into a multitransgenic pig;
• wherein the porcine cell and the multitransgenic pig lack expression of alpha 1,3 galactosyltransferase (GTKO);
• and
• wherein the transgenic porcine animal comprises a deletion of a gene selected from the group consisting of CMAH, B4GalNT2, GGTA1, GHR, and a combination thereof.

A transgenic porcine animal comprising genetic modifications that result in:

• (i) the lack of expression of functional alpha 1,3 galactosyltransferase;
• and
• (ii)
• incorporation and expression at a single genomic locus of a polycistronic vector comprising SEQ ID NO:
• 12.