Claims
- 1. A method of stimulating site-specific DNA recombination in a cell that is genetically engineered to undergo site-specific DNA recombination mediated by a site-specific DNA recombinase, comprising contacting the cell with a polypeptide comprising the site-specific DNA recombinase and a membrane translocation sequence, thereby stimulating site-specific DNA recombination in the cell.
- 2. A method of stimulating site-specific DNA recombination in an animal, comprising administering a polypeptide comprising a site-specific DNA recombinase and a membrane translocation sequence to an animal comprising a cell that is genetically engineered to undergo site-specific recombination mediated by the site-specific DNA recombinase, thereby stimulating site-specific DNA recombination in the animal.
- 3. A method of identifying a compound that modulates the delivery of a polypeptide to a cell or the activity of a polypeptide in a cell, comprising:
a) contacting a population of cells with the compound, wherein the population comprises cells that are genetically engineered to under site-specific recombination, b) contacting the population of cells with a polypeptide comprising a site-specific DNA recombinase and a membrane translocation sequence; and c) detecting site-specific recombination mediated by the site-specific DNA site-specific recombinase, whereby an increase or decrease in the number of cells that undergo site-specific recombination, compared to the number of cells that undergo site-specific recombination in a population of cells not contacted by the compound, identifies a compound that modulates the delivery of a polypeptide to a cell or the activity of a polypeptide in a cell.
- 4. A method of identifying an amino acid sequence that modulates the delivery of a polypeptide to a cell or the activity of a polypeptide in a cell, comprising:
a) contacting a population of cells with a polypeptide comprising a site-specific DNA recombinase and a membrane translocation sequence and an additional amino acid sequence, wherein the population comprises cells that are genetically engineered to undergo site-specific DNA recombination; and b) detecting site-specific recombination mediated by the site-specific DNA site-specific recombinase, whereby an increase or decrease in the number of cells that undergo site-specific recombination induced by the polypeptide comprising the site-specific DNA recombinase, the membrane translocation sequence, and the additional amino acid sequence, compared to the number of cells that undergo site-specific recombination induced by a polypeptide comprising the site-specific DNA recombinase and the membrane translocation sequence and lacking the additional amino acid sequence, identifies an amino acid sequence that modulates the delivery of a polypeptide to a cell or the activity of a polypeptide in a cell.
- 5. The method of claim 1, 2, 3, or 4, wherein the site-specific DNA recombinase is Cre recombinase.
- 6. The method of claim 1, 2, 3, or 4, wherein the site-specific DNA recombinase is Flp recombinase.
- 7. The method of claim 1, 2, 3, or 4, wherein the polypeptide further comprises a nuclear localization sequence.
- 8. An isolated polypeptide comprising a site-specific DNA recombinase and a membrane translocation sequence.
- 9. The isolated polypeptide of claim 8, wherein the isolated polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 1.
- 10. An isolated nucleic acid encoding a polypeptide comprising a site-specific DNA recombinase and a membrane translocation sequence.
- 11. The isolated nucleic acid of claim 10, wherein the isolated nucleic acid encodes the amino acid sequence set forth in SEQ ID NO: 1.
- 12. The isolated nucleic acid of claim 10, wherein the isolated nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO: 2.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit of priority to U.S. Ser. No. 60/230,690, filed Sep. 7, 2000.
STATEMENT OF FEDERALLY SPONSORED RESEARCH
[0002] This invention was made with government support under Grant No. R01RR13166 awarded by the Public Health Service, National Institutes of Health. The government has certain rights in the invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60230690 |
Sep 2000 |
US |