Combinational usage of insulators and coat color marker for high efficiency transgene expression and visual genotyping in mice
AssigneeLevel Biotechnology Inc.
InventorsTing-Fen Tsai, Yi-Chun Hsiao, Hsiao-Hui Chang
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Abstract
A method of identifying a genotype of a transgenic mouse and reducing the variability of a transagene expression in the transgenic mouse comprises the step of introducing into a mouse (a) a transgene expression cassette, (b) a coat color expression cassette and (c) an insulator positioned at 5′ or 3′ end of the transgene expression cassette.
Claims14
A method of producing a transgenic mouse comprising:
- introducing a vector into a mouse embryo or a mouse ES cell and transferring said ES cell into a blastocyst;
- transferring said embryo or said blastocyst comprising said ES cell into a pseudopregnant female mouse;
- allowing said embryo or blastocyst to develop into an offspring;
- selecting an offspring that expresses agouti cDNA and has a coat color phenotype;
- wherein said vector comprises:
- a first transgene expression cassette comprising mouse agouti cDNA operably linked to a human keratinocyte specific K14 promoter, a second transgene expression cassette comprising RNA polymerase II large subunit promoter, and a chicken beta-globin HS4 insulator;
- wherein said insulator and said first transgene expression cassette are located at the 5′ or 3′ end of said second transgene expression cassette;
- the number of copies of said chicken beta-globin HS4 insulator is 1–6;
- and said insulator is in the same or opposite orientation relative to said first and second transgene expression cassettes in said vector.
A vector comprising a first transgene expression cassette comprising mouse agouti cDNA operably linked to a human keratinocyte specific K14 promoter, a second transgene expression cassette comprising RNA polymerase II large subunit promoter, and a chicken beta-globin HS4 insulator;
- wherein said insulator and said first transgene expression cassette are located at the 5′ or 3′ end of said second transgene expression cassette;
- the number of copies of said chicken beta-globin HS4 insulator is 1–6 and said insulator is in the same or opposite orientation relative to said first and second transgene expression cassettes in said vector.
A method of producing a transgenic mouse comprising:
- introducing a vector into a mouse embryo or a mouse ES cell and transferring said ES cell into a blastocyst;
- transferring said embryo or said blastocyst comprising said ES cell into a pseudopregnant female mouse;
- allowing said embryo or blastocyst to develop into an offspring;
- selecting an offspring that expresses agouti cDNA and has a coat color phenotype;
- wherein said vector comprises:
- a first transgene expression cassette comprising mouse agouti cDNA operably linked to a promoter, a second transgene expression cassette comprising RNA polymerase II large subunit promoter, and a chicken beta-globin HS4 insulator.
A vector comprising a first transgene expression cassette comprising mouse agouti cDNA operably linked to a promoter, a second transgene expression cassette comprising RNA polymerase II large subunit promoter, and a chicken beta-globin HS4 insulator.
Dependencies are inferred from the claim text.