US2008261277A1PendingUtilityA1

pRNA chimera

Assignee: PURDUE RESEARCH FOUNDATIONPriority: Aug 23, 2000Filed: Jun 2, 2008Published: Oct 23, 2008
Est. expiryAug 23, 2020(expired)· nominal 20-yr term from priority
C12N 2320/32C12N 15/1131C12N 2310/53C12N 15/111C12N 15/1137C12N 2310/111C12N 2310/121C12N 2310/3519C12N 2310/11A61K 38/00C12N 2310/16A61P 31/00C12N 15/113A61P 35/00C12Y 113/11031C12N 2799/022
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Claims

Abstract

A circularly permuted chimeric pRNA molecule carrying a stabilized biologically active RNA, such as a ribozyme.

Claims

exact text as granted — not AI-modified
1 . A pRNA chimera comprising a small interfering RNA (siRNA). 
     
     
         2 . The pRNA chimera of  claim 1  comprising a base-paired region comprising the siRNA. 
     
     
         3 . The pRNA chimera of  claim 1  which is circularly permuted. 
     
     
         4 . The pRNA chimera of  claim 3  comprising a pRNA region and a spacer region, wherein the pRNA region comprises a based-paired region comprising a first biologically active nucleic acid comprising the siRNA, and the spacer region comprises a second biologically active nucleic acid. 
     
     
         5 . The pRNA chimera of  claim 4  wherein the second biologically active nucleic acid comprises a therapeutic agent. 
     
     
         6 . The pRNA chimera of  claim 4  wherein the second biological active nucleic acid possesses enzymatic activity or binding activity or both. 
     
     
         7 . The pRNA chimera of  claim 4  wherein the second biologically active nucleic acid is selected from the group consisting of a ribozyme, an RNA aptamer, an antisense RNA and a peptide nucleic acid (PNA). 
     
     
         8 . The pRNA chimera of  claim 1  which is not circularly permuted. 
     
     
         9 . The pRNA chimera of  claim 1  comprising a 5′ or 3′ end comprising an end-labeling agent. 
     
     
         10 . The pRNA chimera of  claim 9  wherein the end-labeling agent is selected from the group consisting of biotin, pCp, DIG, SH group and phosphate. 
     
     
         11 . The pRNA chimera of  claim 1  comprising a 5′ or 3′ end comprising an exogenous RNA. 
     
     
         12 . The pRNA chimera of  claim 1  comprising a 5′ or 3′ end comprising an RNA aptamer. 
     
     
         13 . The pRNA chimera of  claim 1  comprising a 5′ or 3′ end comprising a therapeutic agent. 
     
     
         14 . The pRNA chimera of  claim 1  comprising at least one nucleotide analog or modified nucleotide. 
     
     
         15 . The pRNA chimera of  claim 1  wherein the nucleotide analog or modified nucleotide is selected from the group consisting of a DNA phosphorothoiate, an RNA phosphorothoiate and a 2′-O-methyl ribonucleotide. 
     
     
         16 . A DNA molecule comprising a nucleotide sequence that encodes a pRNA chimera comprising an siRNA. 
     
     
         17 . A method for making a pRNA chimera comprising:
 providing a DNA encoding a pRNA chimera comprising an siRNA; and   transcribing the DNA in vitro to yield the pRNA chimera.   
     
     
         18 . The method of  claim 17  further comprising using polymerase chain reaction on a DNA template to generate the DNA encoding the pRNA chimera. 
     
     
         19 . The method of  claim 17  further comprising generating the DNA encoding the pRNA chimera by cloning the DNA into a plasmid and replicating the plasmid. 
     
     
         20 . A method for delivering siRNA to a cell comprising:
 introducing into the cell a DNA molecule comprising a nucleotide sequence that operably encodes the pRNA chimera comprising an siRNA; and   causing transcription of the DNA to yield the siRNA.   
     
     
         21 . The method of  claim 20  wherein the pRNA chimera is a circularly permuted pRNA chimera comprising a pRNA region and a spacer region, wherein the pRNA region comprises a based-paired region comprising a first biologically active nucleic acid comprising the siRNA, and the spacer region comprises a second biologically active nucleic acid. 
     
     
         22 . The method of  claim 21  wherein the second biologically active RNA is selected from the group consisting of a ribozyme, an RNA aptamer, an antisense RNA and a peptide nucleic acid (PNA). 
     
     
         23 . The method of  claim 20  wherein the cell is present in a cell culture, a tissue, an organ or an organism. 
     
     
         24 . The method of  claim 20  wherein the cell is a mammalian cell. 
     
     
         25 . The method of  claim 24  wherein the cell is a human cell. 
     
     
         26 . A polyvalent multimeric complex comprising a plurality of chimeric pRNA monomers comprising:
 at least one chimeric pRNA monomer comprising a first biologically active nucleic acid comprising an siRNA; and   at least one circularly permuted chimeric pRNA monomer comprising a pRNA region and a spacer region, wherein the spacer region comprises a second biologically active nucleic acid.   
     
     
         27 . The polyvalent multimeric complex of  claim 26  wherein the second biologically active RNA is selected from the group consisting of a ribozyme, an RNA aptamer, an antisense RNA and a peptide nucleic acid (PNA). 
     
     
         28 . The polyvalent multimeric complex of  claim 26  wherein the second biologically active RNA comprises an anti-receptor RNA. 
     
     
         29 . The polyvalent multimeric complex of  claim 26  wherein at least one chimeric pRNA monomer comprises a 5′ or 3′ end comprising an end-labeling agent. 
     
     
         30 . The polyvalent multimeric complex of  claim 29  wherein the end-labeling agent is selected from the group consisting of biotin, pCp, DIG, SH group and phosphate. 
     
     
         31 . The polyvalent multimeric complex of  claim 26  wherein at least one chimeric pRNA monomer comprises a 5′ or 3′ end comprising an exogenous RNA. 
     
     
         32 . The polyvalent multimeric complex of  claim 26  wherein at least one chimeric pRNA monomer comprises a 5′ or 3′ end comprising an RNA aptamer. 
     
     
         33 . The polyvalent multimeric complex of  claim 26  wherein at least one chimeric pRNA monomer comprises a 5′ or 3′ end comprising a therapeutic agent. 
     
     
         34 . The polyvalent multimeric complex of  claim 26  wherein at least one chimeric pRNA monomer comprises at least one nucleotide analog or modified nucleotide. 
     
     
         35 . The polyvalent multimeric complex of  claim 34  wherein the nucleotide analog or modified nucleotide is selected from the group consisting of a DNA phosphorothoiate, an RNA phosphorothoiate and a 2′-O-methyl ribonucleotide. 
     
     
         36 . The polyvalent multimeric complex of  claim 26  which is a pRNA hexamer.

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