EP0321201A2 — Ribozymes — Luxuse Patents

Claim 1

Independent Claim 41 words

A ribozyme comprising a hybridizing region which is complementary in nucleotide sequence to at least part of a target RNA, and a catalytic region which is adapted to cleave said target RNA wherein the hybridizing region contains 9 or more nucleotides.

Claim 2

Dependent Claim 41 words

A ribozyme comprising a hybridizing region which is complementary in nucleotide sequence to at least part of a target RNA, and a catalytic region which is adapted to cleave said target RNA wherein the hybridizing region contains 9 or more nucleotides.

Claim 3

Dependent Claim 84 words

A ribozyme comprising one or more arms formed of single stranded RNA and having a sequence complementary to at least part of a target RNA, said one or more arms being associated with a catalytic region, capable of cleaving said target RNA; and where the hybridizing region comprises a single arm of RNA said arm contains at least 9 nucleotides, and where the hybridizing region comprises 2 or more arms of RNA, the sum of nucleotides in said arms is greater than 9 nucleotides.

Claim 4

Dependent Claim 84 words

A ribozyme comprising one or more arms formed of single stranded RNA and having a sequence complementary to at least part of a target RNA, said one or more arms being associated with a catalytic region, capable of cleaving said target RNA; and where the hybridizing region comprises a single arm of RNA said arm contains at least 9 nucleotides, and where the hybridizing region comprises 2 or more arms of RNA, the sum of nucleotides in said arms is greater than 9 nucleotides.

Claim 5

Dependent Claim 21 words

A ribozyme according to claim 2, having two single stranded arms of RNA, associated with one another through said catalytic region.

Claim 6

Dependent Claim 14 words

A ribozyme according to any preceding claim which does not modify cleaved target RNA.

Claim 7

Dependent Claim 117 words

A ribozyme of the formula: wherein; X represents any ribonucleotide and each X residue may be the same or different; the sum of n and n′ is greater than 6 and n and n′ may be the same or different; an (*) represents a base pair between complementary ribonucleotides; X′ and X˝ represent ribonucleotides of complementary sequence along at least part of their length to allow base pairing between the oligoribonucleotides, or X′ and X˝ together form a single RNA sequence wherein at least part of said sequence comprises a stem formed by base pairing between complementary nucleotides; and optionally, an additional nucleotide selected from any one of A, G, C or U is inserted after ¹A.

Claim 8

Dependent Claim 117 words

A ribozyme of the formula: wherein; X represents any ribonucleotide and each X residue may be the same or different; the sum of n and n′ is greater than 6 and n and n′ may be the same or different; an (*) represents a base pair between complementary ribonucleotides; X′ and X˝ represent ribonucleotides of complementary sequence along at least part of their length to allow base pairing between the oligoribonucleotides, or X′ and X˝ together form a single RNA sequence wherein at least part of said sequence comprises a stem formed by base pairing between complementary nucleotides; and optionally, an additional nucleotide selected from any one of A, G, C or U is inserted after ¹A.

Claim 9

Dependent Claim 92 words

A ribozyme as claimed in any preceding claim having the formula: wherein; X represents any ribonucleotide and each X residue may be the same or different; an (*) represents a base pair between complementary ribonucleotides; n and n′ are as previously defined; B represents a bond, a base pair, a ribonucleotide, or an oligoribonucleotide containing at least 2 ribonucleotides; m and m′ are 1 or more and may be the same or different; and optionally, an additional nucleotide selected from any one of A, G, C or U is inserted after ¹A.

Claim 10

Dependent Claim 22 words

A ribozyme according to any one of claims 5 or 6, where the sum of n and n′ is 16 or greater.

Claim 11

Dependent Claim 40 words

A ribozyme according to any preceding claim wherein the catalytic region is capable of cleaving a target RNA after the sequence X o UY, where X o is any ribonucleotide, U is uracil and Y is adenine, cytosine or uracil.

Claim 12

Dependent Claim 27 words

A ribozyme containing multiple catalytic domains connected through flanking sequences, each said flanking sequence including sequences complementary to the target RNA which is desired to be inactivated.

Claim 13

Dependent Claim 27 words

A ribozyme containing multiple catalytic domains connected through flanking sequences, each said flanking sequence including sequences complementary to the target RNA which is desired to be inactivated.

Claim 14

Dependent Claim 13 words

A ribozyme according to any preceding claim which has been stabilized by circularization.

Claim 15

Dependent Claim 12 words

A ribozyme comprised of catalytic antisense sequences according to any preceding claim.

Claim 16

Independent Claim 30 words

A method for the inactivation or cleavage of a target RNA which comprises reacting said target RNA with a ribozyme as claimed in any one of claims 1 to 11.

Claim 17

Dependent Claim 66 words

A method according to claim 12, wherein said inactivation occurs in-vivo in prokaryotic or eukaryotic cells, as a result of transcription of said ribozyme from a DNA or RNA transfer vector transfected into said cells or precursors thereof, or expression of a gene encoding said ribozyme which is integrated into the host genome, such that said ribozyme hybridizes to and inactivates a desired cellular target RNA.

Claim 18

Dependent Claim 14 words

A method according to claim 13, wherein said cells are plant or animal cells.

Claim 19

Dependent Claim 24 words

A method according to claim 13 or 14, wherein the target RNA is endogenous to the cells, or is exogenous, such as viral RNA.

Claim 20

Dependent Claim 25 words

A method according to claim 15, wherein target viral RNA is selected from RNA of the HIV(AIDS), hepatitis or other virus causing disease in mammals.

Claim 21

Dependent Claim 13 words

A method according to claim 15, wherein the viral RNA is bacteriophage RNA.

Claim 22

Dependent Claim 43 words

Use of a ribozyme according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of viral disease in man or animals, said ribozyme being capable of hybridizing to and inactivating transcripts of said viral RNA.

Claim 23

Independent Claim 38 words

A method for treating viral or viroid disease in plants which comprises administering to a plant a ribozyme according to any one of claims 1 to 11, said ribozyme hybridizing to and inactivating transcripts of said viral RNA.

Claim 24

Dependent Claim 27 words

A composition which comprises a ribozyme as claimed in any one of claims 1 to 11 in association with pharmaceutically, veterinarially, or agriculturally acceptable carrier or excipient.

Claim 25

Independent Claim 58 words

A method for producing a ribozyme according to any one of claims 1 to 11 which comprises the steps of: (a) ligating into a transfer vector comprised of DNA, RNA or a combination thereof a nucleotide sequence corresponding to said ribozyme; (b) transcribing the nucleotide sequence of step (a) with RNA polymerase; and (c) optionally purifying the ribozyme.

Claim 26

Dependent Claim 34 words

A transfer vector comprised of RNA or DNA or a combination thereof containing a nucleotide sequence which on transcription gives rise to a ribozyme as claimed in any one of claims 1 to 11.

Claim 27

Dependent Claim 15 words

A transfer vector according to claim 22, which is a bacterial plasmid or phage DNA.

Claim 28

Dependent Claim 27 words

A prokaryotic or eukaryotic cell containing nucleotide sequences which on transcription thereof give rise to a ribozyme as claimed in any one of claims 1 to 11.

Ribozymes

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