Covalent lock for self-assembled oligonucleotide constructs

• US 5,780,613 A
• Filed: 08/01/1995
• Issued: 07/14/1998
• Est. Priority Date: 08/01/1995
• Status: Expired due to Fees

• Current Assignee: Northwestern University
• Original Assignee: Northwestern University
• Inventors: Herrlein, Mathias K., Letsinger, Robert L.
• Primary Examiner(s): ARTHUR, LISA BENNETT
• Application Number:
  US08/509,858
• Time in Patent Office: 1,078 Days
• Field of Search: 435/6, 536/22.1, 536/23.1, 536/25.3, 536/25.33
• US Class Current: 536/25.33
• 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

Abstract

A method of autoligating self-assembled oligonucleotide blocks is disclosed. The method includes the step of displacing a 5'"'"' displaceable group by a 3'"'"' thiophosphoryl group to form an --OP(O)(O^-)S-- internucleoside linkage.

Claims

1. A method of covalently autoligating self-assembled oligonucleotide segments by displacing a 5'"'"' displaceable group by a 3'"'"' thiophosphoryl group to spontaneously form an --OP (O) (O³¹ ) S-- internucleoside linkage.

2. A method of claim 1 further including the steps of bringing the 5'"'"' displaceable group and 3'"'"'-thiophosphoryl group into proximity, the 3'"'"' thiophosphoryl group then displacing the displaceable group to form the internucleoside linkage.

3. A method as set forth in claim 2 wherein the 3'"'"'-thiophosphoryl group and the 5'"'"' displaceable group are on the same oligonucleotide.

4. A method as set forth in claim 3 wherein said forming step is further defined as forming a cyclized oligonucleotide containing non-nucleotide bridges.

5. A method as set forth in claim 3 wherein said forming step is further defined as forming a dumbbell oligonucleotide.

6. A method as set forth in claim 3 wherein said forming step is further defined as forming a cyclized nucleotide conjugate.

7. A method as set forth in claim 2 wherein the 3'"'"'-thiophosphoryl group and the 5'"'"' displaceable group are on different oligonucleotide strands.

8. A method as set forth in claim 1 wherein the displaceable group is selected from the group consisting of Cl^-, Br^-,I^-, and RSO₃^- where R is phenyl or phenyl substituted with one to five atoms or groups comprising F,Cl,Br,I,alkyl (C1 to C6), nitro, cyano, sulfonyl and carbonyl, or R is alkyl with one to six carbons.

9. A method as set forth in claim 1 wherein the displaceable group is a hydrophobic tosyl group, said displacing step being further defined as displacing the 5'"'"'-O-tosyl group by a "3'"'"'-thiophosphoryl" group.

10. A method as set forth in claim 9 wherein the tosyl group is a 5'"'"'-O-p-toluenesulfonyl group, said displacing step being further defined as displacing the 5'"'"'-O-p-toluenesulfonyl group by the 3'"'"'-thiophosphoryl group.