Tamoxifen and 4-hydroxytamoxifen-activated system for regulated production of proteins in eukaryotic cells
AssigneeThe Board of Trustees of the University of Illinois
InventorsChengjian Mao, David J. Shapiro
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Abstract
Novel tamoxifen inducible and ICI 182,780 repressible expression systems comprising mutant estrogen receptors and mutant estrogen response element are disclosed. Such systems have a wide variety of applications, including gene therapy and in vivo and in vitro expression, as well as their use in transgenic animals.
Claims51
An isolated polynucleotide selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 75, 78, 80 and 82, and complements thereof.
An isolated polynucleotide comprising a nucleotide sequence having at least more than 80% sequence identity to the sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 75, 78, 80 and 82, and complements thereof.
A vector, comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21 and 23, and complements thereof.
A vector, comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82.
A vector, comprising:
- at least one copy of a first polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21 and 23, and complements thereof, and
- at least one copy of a second polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82.
An isolated polynucleotide comprising a first polynucleotide selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, operably linked to a second polynucleotide.
An isolated polypeptide encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21 and 23.
An isolated polypeptide selected from the group consisting of SEQ ID NOS:
- 4, 6, 8, 10, 12, 14, 16, 18, 22 or 24.
An isolated polypeptide having at least 80% sequence identity to a polypeptide sequence selected from the group consisting of SEQ ID NOS:
- 4, 6, 8, 10, 12, 14, 16, 18, 22 and 24.
An isolated polypeptide comprising:
- a first polypeptide comprising a VP16 domain, fused to a second polypeptide selected from the group consisting of SEQ ID NO:
- 4, 6, 8, 10, 12, 14, 16, 18, 22 and 24.
An isolated polynucleotide having a nucleotide sequence comprising SEQ. ID NOS:
- 66, 68, 70 or 72.
An isolated polypeptide encoded by a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 66, 68, 70 and 72.
An isolated polynucleotide having at least 80% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOS:
- 66, 68, 70 and 72.
An isolated polypeptide selected from the group consisting of SEQ ID NOS:
- 67, 69, 71 and 73.
An isolated polypeptide having at least 80% sequence identity to a polypeptide selected from the group consisting of SEQ ID NOS:
- 67, 69, 71 and 73.
An inducible expression system, comprising:
- a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72, and complements thereof, and
- a second polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, and complements thereof.
A kit, comprising:
- at least one copy of a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72, and complements thereof;
- and
- at least one copy of a second polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, and complements thereof.
A method of inducing gene expression in a subject comprising:
- administering to the subject at least one copy of a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72 and complements thereof, and
- administering to the subject at least one copy of a second polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, and complements thereof, that is operably linked to the gene;
- such that upon induction with an inducing agent, the gene is expressed.
A method of silencing gene expression in a subject, comprising:
- administering to the subject at least one copy of a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72 and complements thereof;
- administering to the subject at least one copy of a second polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, and complements thereof, that is operably linked to the gene;
- such that upon induction with an inducing agent, the gene is expressed;
- and
- administering a repressing agent.
A method of inhibiting endogenous gene expression in a subject, comprising:
- administering to the subject a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72, and complements thereof;
- and
- administering to the subject a second polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82 and complements thereof,
- wherein said second polynucleotide is operably linked to a third polynucleotide,
- such that upon induction with an inducing agent, said third polynucleotide is transcribed in an anti-sense orientation to the sequence of the endogenous gene.
A non-human transgenic animal, comprising:
- a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72, and complements thereof;
- and
- a second polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, and complements thereof.
A cell, comprising
- a first polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 66, 68, 70 and 72, and complements thereof;
- and
- a second polynucleotide comprising a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS:
- 75, 78, 80 and 82, and complements thereof.
Dependencies are inferred from the claim text.