Oligonucleotides with selectably cleavable and/or abasic sites

• US 5,367,066 A
• Filed: 07/24/1991
• Issued: 11/22/1994
• Est. Priority Date: 10/16/1984
• Status: Expired due to Fees

• Current Assignee: Chiron Diagnostics Corp (Bayer AG)
• Original Assignee: Chiron Corporation (Novartis Ag)
• Inventors: Urdea, Michael S., Horn, Thomas
• Primary Examiner(s): Brown, Johnnie R.
• Assistant Examiner(s): Kunz, Gary L.
• Application Number:
  US07/736,445
• Time in Patent Office: 1,217 Days
• Field of Search: 536/27, 536/23.1, 536/24.3, 536/24.31, 536/24.32, 435/6
• US Class Current: 536/24.3
• CPC Class Codes: C07F 9/10 Phosphatides, e.g. lecithin
• CPC Class Codes: C07F 9/2429 of arylalkanols
• CPC Class Codes: C07F 9/65515 the oxygen atom being part of a five-membered ring
• CPC Class Codes: C07H 15/18 Acyclic radicals, substituted by carbocyclic rings
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6823 Release of bound markers
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/683 involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
• CPC Class Codes: C12Q 2521/301 Endonuclease

Abstract

A modified polynucleotide containing at least one cleavable or abasic site as shown below. ##STR1## DNA₁ is a first segment of DNA; DNA₂ is a second segment of DNA; and R_m is C₁ to C₁₆ alkylene or an oxytheylene oligomer --(CH₂ CH₂ O)_z -- where z is an interger in the range of 1 to 16 inclusive, and R_n is selected from the group consisting of ##STR2## Such polynucleotides are useful in solid phase hybridizations because they permit the release of a label from the solid support after the hybridization reaction.

Claims

1. A polynucleotide reagent having the structure ##STR30## wherein DNA₁ is a first segment of DNA;

DNA₂ is a second segment of DNA; and

R_m is C₁ -C₁₆ alkylene or an oxyethylene oligomer --(CH₂ CH₂ O)_z -- where z is an integer in the range of 1 to 16 inclusive, and R_n is selected from the group consisting of ##STR31##

2. The polynucleotide reagent of claim 1, wherein R_m is --(CH₂ CH₂ O)₄ -- and R_n is ##STR32##

3. The polynucleotide reagent of claim 2, wherein R_m is C₁ -C₁₆ alkylene.

4. The polynucleotide reagent of claim 1, wherein R_m is C₂ -C₁₂ alkylene.

5. The polynucleotide reagent of claim 1, wherein R_m is an oxyethylene oligomer --(CH₂ CH₂ O)_z -- where z is an integer in the range of 2 to 12 inclusive.

6. The polynucleotide reagent of claim 1, wherein R_n is ##STR33##

7. The polynucleotide reagent of claim 4, wherein R_n is ##STR34##

8. The polynucleotide reagent of claim 5, wherein R_n is ##STR35##

9. The polynucleotide reagent of claim 1, wherein R_n is ##STR36##

10. The polynucleotide reagent of claim 4, wherein R_n is ##STR37##

11. The polynucleotide reagent of claim 5, wherein R_n is ##STR38##

12. The polynucleotide reagent of claim 1, wherein R_n is ##STR39##

13. The polynucleotide reagent of claim 4, wherein R_n is ##STR40##

14. The polynucleotide reagent of claim 5, wherein R_n is ##STR41##

15. The polynucleotide reagent of claim 1, wherein R_n is ##STR42##

16. The polynucleotide reagent of claim 4, wherein R_n is ##STR43##

17. The polynucleotide reagent of claim 5, wherein R_n is ##STR44##

18. The polynucleotide reagent of claim 1, wherein R_n is ##STR45##

19. The polynucleotide reagent of claim 4, wherein R_n is ##STR46##

20. The polynucleotide reagent of claim 5, wherein R_n is ##STR47##

21. The polynucleotide reagent of claim 1, wherein R_n is

22. The polynucleotide reagent of claim 4, wherein R_n is

23. The polynucleotide reagent of claim 5, wherein R_n is