Telomerase reverse transcriptase transcriptional regulatory sequences and methods of using
AssigneeGeron Corporation
InventorsGregg B. Morin, Serge P. Lichtsteiner, Alain P. Vasserot, Robert R. Adams, William H. Andrews
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Abstract
The present invention is related to novel nucleic acids comprising telomerase reverse transcriptase (TERT) cis-acting transcriptional control sequences, including TERT human and mouse promoter sequences. The present invention is further directed to methods of using these cis-acting transcriptional control sequences, for example, to drive heterologous gene sequences; to modulate the level of transcription of TERT or to isolate novel trans-acting regulatory factors which bind to and modulate the activity of a TERT promoter.
Claims20
A method of killing a mammalian cell that expresses telomerase reverse transcriptase (TERT), comprising contacting the cell with a polynucleotide in which a promoter sequence controls transcription of a transcribable sequence that is toxic to the cell or renders the cell more susceptible to toxicity of a drug;
- wherein the promoter has the property of causing the transcribable sequence to be expressed in cells endogenously expressing TERT, and contains a nucleotide sequence that is least 90% identical to the sequence from position −117 to position −36 from the translation initiation site (position 13545) of SEQ. ID NO:1.
A method of killing a mammalian cell that expresses telomerase reverse transcriptase (TERT), comprising contacting the cell with a polynucleotide in which a promoter sequence controls transcription of a transcribable sequence that is toxic to the cell or renders the cell more susceptible to toxicity of a drug;
- wherein the promoter has the property of causing the transcribable sequence to be expressed in cells endogenously expressing TERT, and is either;
- a) contained in the APAI-FSPI fragment just upstream of the encoding sequence for human telomerase reverse transcriptase (hTERT) in lambda phage GΦ5 deposited as ATCC Accession No. 98505;
- or
- b) comprises a nucleotide sequence that hybridizes to DNA complementary to said APAI-FSPI fragment at 5 to 10° C. below Tm in aqueous solution at 1 M NaCl followed by wash in 0.2×SSC, wherein Tm is the melting temperature of the APAI-FSPI fragment in double-stranded form.
A method of treating cancer in a subject, comprising contacting cancer cells in the subject with a polynucleotide in which a promoter sequence controls transcription of a transcribable sequence that is toxic to the cell or renders the cell more susceptible to toxicity of a drug;
- wherein the promoter has the property of causing the transcribable sequence to be expressed in cells endogenously expressing TERT, and contains a nucleotide sequence that is least 90% identical to the sequence from position −117 to position −36 from the translation initiation site (position 13545) of SEQ. ID NO:1.
A method of expressing a transcribable nucleotide sequence in a cell, comprising contacting the cell with a polynucleotide in which the transcribable nucleotide sequence is operably linked to a promoter sequence so as to cause it to be transcribed when the polynucleotide is in cells endogenously expressing human telomerase reverse transcriptase (hTERT);
- wherein the promoter has the property of causing the transcribable sequence to be expressed in cells endogenously expressing TERT, and contains a nucleotide sequence that is least 90% identical to the sequence from position −117 to position −36 from the translation initiation site (position 13545) of SEQ. ID NO:1.
A polynucleotide in which a promoter is operably linked to a heterologous encoding region so as to cause it to be transcribed when the polynucleotide is in cells endogenously expressing human telomerase reverse transcriptase (hTERT),
- wherein the promoter contains a nucleotide sequence that is least 90% identical to the sequence from position −117 to position −36 from the translation initiation site (position 13545) of SEQ. ID NO:1.
A polynucleotide in which a promoter is operably linked to a heterologous sequence so as to cause the heterologous sequence to be transcribed when the polynucleotide is in cells endogenously expressing human telomerase reverse transcriptase (hTERT),
- wherein the promoter is either
- a) contained in the APAI-FSPI fragment just upstream of the encoding sequence for human telomerase reverse transcriptase (hTERT) in lambda phage GΦ5 deposited as ATCC Accession No. 98505;
- or
- b) comprises a nucleotide sequence that hybridizes to DNA complementary to said APAI-FSPI fragment at 5 to 10° C. below Tm in aqueous solution at 1 M NaCl followed by wash in 0.2×SSC, wherein Tm is the melting temperature of the APAI-FSPI fragment in double-stranded form.
Dependencies are inferred from the claim text.