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US5707835A
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Ribozymes
Assignee: Gene Shears Pty Ltd
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Filing: 1995-05-17
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Granted: 1998-01-13
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Expires: 2015-05-17
Abstract
Abstract Compounds having highly specific endoribonuclease activity are described. The compounds of this invention, also known as ribozymes, comprise ribonucleotides having two hybridizing regions with predetermined sequences capable of hybridizing with a plant, animal or viral target RNA, a region of defined sequence and a base paired stem region.
Inventors
- 👤 James Phillip Haseloff
- 👤 Wayne Lyle Gerlach
- 👤 Philip Anthony Jennings
- 👤 Fiona Helen Cameron
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 19:01
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; A; A01; A01N; A01N57/00; A01N57/10; A01N57/16; A01N63/00; A01N63/50; C12N15/63; C12N15/79; C12N15/82; C12N15/8216; C12N15/8218; A61; A61K; A61K38/00; C12N2310/00; C12N2310/10; C12N2310/12; C12N2310/121; C12N2310/124
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
345 words
A compound having the formula: 3'-- --(Y).sub.r --Q--(Y).sub.s --!.sub.z --5' wherein each Q represents the structure ##STR10## wherein each X represents a ribonucleotide which may be the same or different; wherein each of (X) n and (X) n' represents an oligoribonucleotide having a predetermined sequence which hybridizes with an RNA target sequence to be cleaved, and (b) does not naturally occur covalently bound to the sequences A--A--A--G--C-- and X--C--U--G--A--, respectively, such RNA target sequence not being present within the compound; wherein each of n and n' represents an integer which defines the number of ribonucleotides in the oligoribonucleotide with the proviso that the sum of n+n' is greater than 6 and sufficient to allow the compound to stably interact with the RNA target sequence through base pairing; wherein each * represents base pairing between the nucleotides located on either side thereof; wherein each solid line represents a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof; wherein a represents an integer which defines a number of ribonucleotides with the proviso that a may be 0 or 1 and if 0, the A located 5' of (X) a is bonded to the G located 3' of (X) a ; wherein each of m and m' represents an integer which is greater than or equal to 1; wherein each of the dashed lines independently represents either a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof or the absence of any such chemical linkage; wherein (X) b represents an oligoribonucleotide which may be present or absent with the proviso that b represents an integer which is greater than or equal to 4 if (X) b is present; wherein one or more of the ribonucleotides may be a ribonucleotide derivative; wherein each Q may independently be the same or different; wherein each Y represents a ribonucleotide which may be the same or different; wherein each of r and s represents an integer greater than or equal to 0; and wherein z represents an integer greater than or equal to 1.
Claim 2
Dependent Claim
345 words
A compound having the formula: 3'-- --(Y).sub.r --Q--(Y).sub.s --!.sub.z --5' wherein each Q represents the structure ##STR10## wherein each X represents a ribonucleotide which may be the same or different; wherein each of (X) n and (X) n' represents an oligoribonucleotide having a predetermined sequence which hybridizes with an RNA target sequence to be cleaved, and (b) does not naturally occur covalently bound to the sequences A--A--A--G--C-- and X--C--U--G--A--, respectively, such RNA target sequence not being present within the compound; wherein each of n and n' represents an integer which defines the number of ribonucleotides in the oligoribonucleotide with the proviso that the sum of n+n' is greater than 6 and sufficient to allow the compound to stably interact with the RNA target sequence through base pairing; wherein each * represents base pairing between the nucleotides located on either side thereof; wherein each solid line represents a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof; wherein a represents an integer which defines a number of ribonucleotides with the proviso that a may be 0 or 1 and if 0, the A located 5' of (X) a is bonded to the G located 3' of (X) a ; wherein each of m and m' represents an integer which is greater than or equal to 1; wherein each of the dashed lines independently represents either a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof or the absence of any such chemical linkage; wherein (X) b represents an oligoribonucleotide which may be present or absent with the proviso that b represents an integer which is greater than or equal to 4 if (X) b is present; wherein one or more of the ribonucleotides may be a ribonucleotide derivative; wherein each Q may independently be the same or different; wherein each Y represents a ribonucleotide which may be the same or different; wherein each of r and s represents an integer greater than or equal to 0; and wherein z represents an integer greater than or equal to 1.
Claim 3
Dependent Claim
27 words
A transfer vector comprised of RNA or DNA or a combination thereof containing a nucleotide sequence which on transcription gives rise to a compound of claim 1.
Claim 4
Dependent Claim
19 words
A transfer vector according to claim 2 which is a bacterial plasmid, viral RNA, viral DNA or phage DNA.
Claim 5
Dependent Claim
40 words
A method of using the transfer vector of claim 2 to produce a compound which comprises transcribing a nucleotide sequence which corresponds to the transfer vector with RNA polymerase under suitable conditions so as to produce and recover said compound.
Claim 6
Dependent Claim
67 words
A method of using the transfer vector of claim 2 which comprises introducing the vector into a suitable eukaryotic host cell and culturing the host cell so as to effect transcription of the nucleotide sequence contained within the transfer vector and thereby produce the compound encoded by such nucleotide sequence, which compound which hybridizes with and specifically cleaving an RNA target sequences within the eukaryotic host cell.
Claim 7
Dependent Claim
52 words
A method of using the transfer vector of claim 2 to produce a compound in a host cell which comprises introducing the transfer vector into a suitable host cell and culturing the host cell under conditions to express the transfer vector so as to thereby produce said compound in the host cell.
Claim 8
Dependent Claim
12 words
The method of claim 6, wherein the host cell is a prokaryote.
Claim 9
Dependent Claim
13 words
The method of claim 6, wherein the host cell is a plant cell.
Claim 10
Dependent Claim
13 words
The method of claim 6, wherein the host cell is an animal cell.
Claim 11
Dependent Claim
21 words
A prokaryotic or cultured eukaryotic cell containing a nucleotide sequence which on transcription gives rise to a compound of claim 1.
Claim 12
Dependent Claim
345 words
A compound having the formula: 3'-- --(Y).sub.r --Q--(Y).sub.s --!.sub.z --5' wherein each Q represents the structure ##STR11## wherein each X represents a ribonucleotide which may be the same or different; wherein each of (X) n and (X) n' represents an oligoribonucleotide having a predetermined sequence which hybridizes with an RNA target sequence to be cleaved, and (b) does not naturally occur covalently bound to the sequences A--A--A--G--C-- and X--C--U--G--A--, respectively, such RNA target sequence not being present within the compound; wherein each of n and n' represents an integer which defines the number of ribonucleotides in the oligoribonucleotide with the proviso that the sum of n+n' is greater than 6 and sufficient to allow the compound to stably interact with the RNA target sequence through base pairing; wherein each * represents base pairing between the nucleotides located on either side thereof; wherein each solid line represents a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof; wherein a represents an integer which defines a number of ribonucleotides with the proviso that a may be 0 or 1 and if 0, the A located 5' of (X) a is bonded to the G located 3' of (X) a ; wherein each of m and m' represents an integer which is greater than or equal to 3; wherein each of the dashed lines independently represents either a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof or the absence of any such chemical linkage; wherein (X) b represents an oligoribonucleotide which may be present or absent with the proviso that b represents an integer which is greater than or equal to 4 if (X) b is present; wherein one or more of the ribonucleotides may be a ribonucleotide derivative; wherein each Q may independently be the same or different; wherein each Y represents a ribonucleotide which may be the same or different; wherein each of r and s represents an integer greater than or equal to 0; and wherein z represents an integer greater than or equal to 1.
Claim 13
Dependent Claim
345 words
A compound having the formula: 3'-- --(Y).sub.r --Q--(Y).sub.s --!.sub.z --5' wherein each Q represents the structure ##STR11## wherein each X represents a ribonucleotide which may be the same or different; wherein each of (X) n and (X) n' represents an oligoribonucleotide having a predetermined sequence which hybridizes with an RNA target sequence to be cleaved, and (b) does not naturally occur covalently bound to the sequences A--A--A--G--C-- and X--C--U--G--A--, respectively, such RNA target sequence not being present within the compound; wherein each of n and n' represents an integer which defines the number of ribonucleotides in the oligoribonucleotide with the proviso that the sum of n+n' is greater than 6 and sufficient to allow the compound to stably interact with the RNA target sequence through base pairing; wherein each * represents base pairing between the nucleotides located on either side thereof; wherein each solid line represents a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof; wherein a represents an integer which defines a number of ribonucleotides with the proviso that a may be 0 or 1 and if 0, the A located 5' of (X) a is bonded to the G located 3' of (X) a ; wherein each of m and m' represents an integer which is greater than or equal to 3; wherein each of the dashed lines independently represents either a chemical linkage providing covalent bonds between the ribonucleotides located on either side thereof or the absence of any such chemical linkage; wherein (X) b represents an oligoribonucleotide which may be present or absent with the proviso that b represents an integer which is greater than or equal to 4 if (X) b is present; wherein one or more of the ribonucleotides may be a ribonucleotide derivative; wherein each Q may independently be the same or different; wherein each Y represents a ribonucleotide which may be the same or different; wherein each of r and s represents an integer greater than or equal to 0; and wherein z represents an integer greater than or equal to 1.
References Cited (Backward Citations)
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