Mammalian telomerase

• US 5,583,016 A
• Filed: 10/27/1994
• Issued: 12/10/1996
• Est. Priority Date: 07/07/1994
• Status: Expired due to Term

• Current Assignee: Geron Corporation
• Original Assignee: Geron Corporation
• Inventors: Andrews, William H., Funk, Walter, Feng, Junli, Villeponteau, Bryant
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Bugaisky, Gabriele E.
• Application Number:
  US08/330,123
• Time in Patent Office: 775 Days
• Field of Search: 536/23.1, 536/23.2, 536/24.31, 536/24.33, 435/194, 435/240.1, 435/252.33, 435/320.1, 435/91.3, 435/91.31, 435/172.3, 435/254.11, 435/252.3
• US Class Current: 435/91.3
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/121 Hammerhead
• CPC Class Codes: C12N 2310/13 Decoys
• CPC Class Codes: C12N 2310/15 Nucleic acids forming more than 2 strands, e.g. TFOs
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 9/1241 Nucleotidyltransferases (2.7.7)
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Y 207/07049 RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase

Abstract

Nucleic acids comprising the RNA component of a mammalian telomerase are useful as pharmaceutical, therapeutic, and diagnostic reagents.

Claims

1. An isolated and purified recombinant nucleic acid fragment comprising an oligonucleotide having a contiguous sequence of at least 25 nucleotides in a sequence complementary or identical to a human genomic DNA sequence encoding the RNA component of human telomerase located in an ˜

2.5 kb HindIII-SacI insert of plasmid pGRN33 (ATCC

     75926).

2. The isolated and purified recombinant nucleic acid of claim 1 that is an oligodeoxyribonucleotide.

3. The isolated and purified recombinant nucleic acid of claim 1 that is an oligoribonucleotide.

4. The isolated and purified recombinant nucleic acid of claim 1 that is complementary to the DNA encoding the RNA component of human telomerase.

5. The isolated and purified recombinant nucleic acid of claim 1 that is plasmid pGRN33 (ATCC 75926).

6. The isolated and purified recombinant nucleic acid of claim 1 that is lambda clone 28-1 (ATCC 75925).

7. The isolated and purified recombinant nucleic acid of claim 1 comprising the sequence:

##STR8## (SEQ ID NO;

     3), wherein "T" also can be "U".

8. The isolated and purified recombinant nucleic acid of claim 1 comprising the sequence:

##STR9## (SEQ ID NO;

     3) wherein "T" also can be "U".

9. The isolated and purified recombinant nucleic acid of claim 3 comprising the sequence:

##STR10## (SEQ ID NO;

     1).

10. The isolated and purified recombinant nucleic acid of claim 1 further comprising a promoter positioned to drive transcription of an RNA complementary in sequence to said oligonucleotide.

11. The isolated and purified recombinant nucleic acid of claim 10 wherein the contiguous sequence is at least 50 nucleotides in length.

12. The isolated and purified recombinant nucleic acid of claim 10 wherein the contiguous sequence is at least 200 nucleotides in length.

13. The isolated and purified recombinant nucleic acid of claim 10 wherein the contiguous sequence is at least 400 nucleotides in length.

14. The isolated and purified recombinant nucleic acid of claim 10 wherein the contiguous sequence is at most 50 nucleotides in length.

15. The isolated and purified recombinant nucleic acid of claim 10 wherein said recombinant nucleic acid functions to produce the oligonucleotide in a prokaryotic host cell.

16. The isolated and purified recombinant nucleic acid of claim 10 wherein said recombinant nucleic acid functions to produce the oligonucleotide in a eukaryotic host cell.

17. The isolated and purified recombinant nucleic acid of claim 16 wherein said recombinant nucleic acid functions to produce the oligonucleotide in a human cell such that the RNA is capable of being assembled by the cell into a functional telomerase molecule.

18. The isolated and purified recombinant nucleic acid of claim 10 wherein the oligonucleotide comprises a human gene for the RNA component of human telomerase.

19. The isolated and purified recombinant nucleic acid of claim 10 comprising the sequence:

##STR11## (SEQ ID NO;

     3), wherein "T" also can be "U".

20. The isolated and purified recombinant nucleic acid of claim 10 comprising the sequence:

##STR12## (SEQ ID NO;

     3) wherein "T" also can be "U".

21. The isolated and purified recombinant nucleic acid of claim 1 comprising a promoter positioned to specifically drive the transcription of an RNA complementary in sequence to said oligonucleotide.

22. An isolated oligonucleotide between 25 and 1000 nucleotides in length in a sequence identical or complementary to a contiguous sequence contained within a human genomic DNA sequence "encoding the RNA component of human telomerase" located in an ˜

2.5 kb HindIII-SacI insert of plasmid pGRN33 (ATCC

     75926).

23. The isolated oligonucleotide of claim 22 at most 200 nucleotides in length.

24. The isolated oligonucleotide of claim 22 at most 50 nucleotides in length.

25. The isolated oligonucleotide of claim 22 at least 50 nucleotides in length.

26. The isolated oligonucleotide of claim 25 at most 200 nucleotides in length.

27. The isolated oligonucleotide of claim 22 at least 200 nucleotides in length.

28. The isolated oligonucleotide of claim 22 at least 400 nucleotides in length.

29. The isolated oligonucleotide of any of claims 22-28 further comprising a label.

30. The isolated oligonucleotide of claim 29 wherein the label is a radioactive label, a fluorescent label, biotin or avidin.

31. Isolated RNA component of human telomerase.

32. An oligonucleotide of SEQ ID NO:

22, 23, 24 or 25.

33. A host cell transformed with a recombinant nucleic acid comprising an oligonucleotide having a contiguous sequence of at least 25 nucleotides in a sequence complementary or identical to a human genomic DNA sequence encoding the RNA component of human telomerase located in an ˜

2.5 kb HindIII-Sac I insert of plasmid pGRN33 (ATCC

     75926).

34. The host cell of claim 33 transformed with pGRN33 (ATCC 75926).

35. The host cell of claim 33 transformed with lambda clone 28-1 (ATCC 75925).

36. The host cell of claim 33 wherein the recombinant nucleic acid further comprises a promoter positioned to drive the transcription of an RNA having a sequence complementary to the oligonucleotide.

37. The host cell of claim 33 that is a eukaryotic cell.

38. The host cell of claim 37 wherein the recombinant nucleic acid encodes an RNA molecule that can associate with protein components of human telomerase to produce telomerase activity capable of adding sequences of repeating units of nucleotides to telomeres.

39. The host cell of claim 38, wherein said repeating unit is 5'"'"'-TTAGGG-3'"'"'.

40. The host cell of claim 38, wherein said repeating unit is not 5'"'"'-TTAGGG-3'"'"'.

41. The host cell of claim 37, wherein said RNA molecule comprises the sequence:

##STR13## (SEQ ID NO;

     1).

42. The host cell of claim 37, wherein the recombinant nucleic acid comprises the sequence:

##STR14## (SEQ ID NO;

3), wherein "T" also can be "U".

43. The host cell of claim 37, wherein the recombinant nucleic acid comprises the sequence:

##STR15## (SEQ ID NO;

     3) wherein "T" also can be "U".

44. A method for producing the RNA component of human telomerase comprising the step of culturing a eukaryotic host cell transformed with a recombinant nucleic acid comprising a promoter positioned to drive the transcription of an oligonucleotide encoding an RNA component of human telomerase.

45. The method of claim 44 wherein the oligonucleotide encoding the RNA component of human telomerase includes a sequence from a human genomic DNA sequence located in an ˜

2.5 kb HindIII-SacI insert of plasmid pGRN33 (ATCC

     75926).

46. The method of claim 44 wherein the cell is a human cell.

47. A method for producing a recombinant telomerase enzyme, said method comprising transforming a eukaryotic host cell capable of expressing protein components of telomerase, with a recombinant nucleic acid comprising a promoter positioned to drive the transcription of an oligonucleotide encoding the RNA component of human telomerase, said recombinant nucleic acid functioning to produce the oligonucleotide in a eukaryotic cell, and culturing said host cells transformed with said vector under conditions such that the protein components and RNA component are expressed and assemble to form an active telomerase molecule capable of adding sequences to telomeres of chromosomal DNA.

48. The method of claim 47 wherein the RNA has a sequence identical to a contiguous sequence encoding the RNA component of human telomerase from a human genomic DNA sequence located in an ˜

2.5 kb HindIII-SacI insert of plasmid pGRN33 (ATCC

     75926).

49. The method of claim 48 wherein the cell is a human cell.