Transgenic pig in which HO-1 and TNFR1-Fc are simultaneously expressed, and method for producing same
The present invention relates to a method for producing a transgenic pig in which immune rejection response is inhibited, and in which human HO-1 genes and TNFR1-Fe fusion genes are simultaneously expressed. The present invention also relates to a transgenic pig for organ transplantation, which is produced by the method, and in which immune rejection response is inhibited. The present invention also relates to a somatic-cell-donating cell strain for producing the transgenic pig, and to a method for producing organs, from the transgenic pig, in which the immune rejection response is inhibited.
1. A method for producing a transgenic pig, comprising:
a) isolating somatic cells from a pig;
b) introducing a gene coding for human HO-1 (Heme oxygenase-1) protein and a gene coding for human TNFR1-Fc (Tumor necrosis factor receptor 1-Fc) fusion protein into the somatic cells;
c) selecting the somatic cells that are introduced with the genes;
d) removing the nucleus from an oocyte of the pig and fusing it with the selected somatic cell to prepare a somatic cell nuclear transferred embryo; and
e) implanting the embryo,
wherein an organ of the transgenic pig is suppressed in immune rejection response during organ xenotransplantation.
2. The method according to claim 1 , wherein the gene coding for human HO-1 protein of step b) has a nucleotide sequence of SEQ ID NO: 1.
3. The method according to claim 1 , wherein the gene coding for human TNFR1-Fc fusion protein of step b) has a nucleotide sequence of SEQ ID NO: 8.
4. The method according to claim 1 , wherein a single vector comprising the gene coding for human HO-1 protein and a single vector comprising the gene coding for human TNFR1-Fc fusion protein are introduced sequentially or simultaneously in step b).
5. The method according to claim 4 , wherein the single vector comprising the gene coding for human HO-1 protein is a vector having a cleavage map of FIG. 1 and the single vector comprising the gene coding for human TNFR1-Fc fusion protein is a vector having a cleavage map of FIG. 4 .
6. The method according to claim 1 , wherein the selected somatic cell in step c) has Accession No. KCLRF-BP-00225.
7. A transgenic pig for organ xenotransplantation, which is produced by the method of claim 1 , wherein an organ of the transgenic pig is suppressed in immune rejection response during organ xenotransplantation.
8. The transgenic pig according to claim 7 , wherein the organ is an islet.
9. A transgenic pig for organ xenotransplantation, which is produced by the method of claim 6 , wherein an organ of the transgenic pig is suppressed in immune rejection response during organ xenotransplantation.
10. The transgenic pig according to claim 9 , wherein the organ is an islet.
11. A somatic donor cell line for the production of the transgenic pig of claim 7 , which is introduced with a gene coding for human HO-1 (Heme oxygenase-1) protein and a gene coding for human TNFR1-Fc (Tumor necrosis factor receptor 1-Fc) fusion protein.
12. The somatic donor cell line according to claim 11 , wherein the somatic donor cell line has Accession No. KCLRF-BP-00225.
13. A method for producing transplantable organs suppressed in immune rejection response, comprising:
producing a transgenic pig by the method of claim 1 ; and
isolating the organ from the transgenic pig.
14. The method according to claim 13 , wherein the organ is an islet.
15. The method according to claim 13 , wherein the organ shows decreased immune rejection response compared to the organ obtained from the pig genetically not modified.
16. A method for producing transplantable organs suppressed in immune rejection response, comprising:
producing a transgenic pig by the method of claim 6 ; and
isolating the organ from the transgenic pig.
17. The method according to claim 16 , wherein the organ is an islet.