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Last Updated: January 26, 2022

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Claims for Patent: 5,965,543

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Summary for Patent: 5,965,543
Title: Senescence responsive transcriptional element
Abstract:Recombinant polynucleotides have expression control sequences that have a senescence responsive element and a minimal promoter, and which are operatively linked to a heterologous nucleotide sequence. The molecules are useful for achieving high levels of expression of genes in senescent cells. Methods of inhibiting expression of genes in senescent cells also are provided.
Inventor(s): Campisi; Judith (Berkeley, CA), Testori; Alessandro (Berkeley, CA)
Assignee: Geron Corporation (Menlo Park, CA)
Application Number:08/840,887
Patent Claims:1. A recombinant polynucleotide comprising an expression control sequence operatively linked to a heterologous nucleotide sequence, the expression control sequence comprising (1) a senescence responsive element comprising a sequence from the interstitial collagenase gene promoter (SEQ ID NO:2), wherein the sequence from the interstitial collagenase gene promoter includes at least 8 consecutive nucleotides from 5'-gaggatgtta taaagcatga gtca-3' (nucleotides 439 to 462 of SEQ ID: NO:2 and terminates on its 3' end no further than nucleotide 478 of SEQ ID NO:2, and (2) a heterologous minimal promoter positioned downstream of the senescence responsive element, wherein the senescence responsive element up-regulates transcription of the heterologous nucleotide sequence in a senescent cell.

2. The recombinant polynucleotide of claim 1 wherein the sequence from the interstitial collagenase gene promoter comprises nucleotides 439 to 462 of the interstitial collagenase gene promoter (5'-gaggatgtta taaagcatga gtca -3' (SEQ ID: NO:2)) and terminates on its 3' end no further than nucleotide 468 of SEQ ID NO:2.

3. The recombinant polynucleotide of claim 2 wherein the sequence from the interstitial collagenase gene promoter comprises nucleotides 428 to 462 of the interstitial collagenase gene promoter (5'-tagctaatcaa gaggatgtta taaagcatga gtca -3' (SEQ ID: NO:2)).

4. The recombinant polynucleotide of claim 2 wherein the sequence comprises nucleotides 394 to 462 of the interstitial collagenase gene promoter (5'-agtc accatttcta atgattgcct agtctattca tagctaatcaa gaggatgtta taaagcatga gtca -3' (SEQ ID: NO:2)).

5. The recombinant polynucleotide of claim 1 wherein the heterologous nucleotide sequence codes for the expression of a transcription regulator, interleukin-6, a tissue inhibitor of a metalloproteinase, collagen, elastin, a growth regulatory transcription factor, an apoptosis regulator or a protein or RNA component of telomerase.

6. The recombinant polynucleotide of claim 1 wherein the heterologous nucleotide sequence codes for the expression of an inhibitor of collagenase, an inhibitor of a metalloproteinase, a plasminogen activator, an inhibitor of a plasminogen activator, interleukin-1 receptor antagonist, an insulin-like growth factors, a cyclin or a cyclin-dependent kinase.

7. The recombinant polynucleotide of any of claims 1, 2, 3 and 4 wherein the sequence terminates on its 5' end no further than nucleotide 394 of SEQ ID NO:2.

8. A polynucleotide probe that specifically hybridizes with a contiguous nucleotide sequence of at least 6 nucleotides within nucleotides 394 to 478SEQ ID NO:2) of the interstitial collagenase gene promoter, or with a contiguous sequence of at least 6 nucleotides complementary to a contiguous nucleotide sequence within nucleotides 394 to 478 (SEQ ID NO:2) of the promoter.

9. The polynucleotide probe of claim 8 further comprising a label.

10. The polynucleotide probe of claim 8 comprising a sequence identical to or complementary to a contiguous nucleotide sequence of at least 6 nucleotides within nucleotides 394 to 478 (SEQ ID NO:2) of the interstitial collagenase gene promoter.

11. The polynucleotide probe of claim 8 comprising a sequence identical to or complementary to a contiguous nucleotide sequence of at least 6 nucleotides within nucleotides 428to 462 (SEQ ID NO:2) of the interstitial collagenase gene promoter.

12. The polynucleotide probe of claim 8 comprising a sequence identical to or complementary to a contiguous nucleotide sequence of at least 6 nucleotides within nucleotides 439 to 462 (SEQ ID NO:2) of the interstitial collagenase gene promoter.

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