Ribozymes

• US 5,254,678 A
• Filed: 08/05/1992
• Issued: 10/19/1993
• Est. Priority Date: 12/15/1987
• Status: Expired due to Term

• Current Assignee: Gene Shears Pty Limited
• Original Assignee: Gene Shears Pty Limited
• Inventors: Haseloff, James P., Jennings, Philip A., Cameron, Fiona H., Gerlach, Wayne L.
• Primary Examiner(s): Schwartz, Richard A.
• Assistant Examiner(s): LEGUYADER, JOHN L
• Application Number:
  US07/926,148
• Time in Patent Office: 440 Days
• Field of Search: 435/91, 435/240.1, 435/172.1, 435/252.3, 435/172.3, 435/320.1, 536/23.1, 536/23.2, 514/44
• US Class Current: 536/23.2
• CPC Class Codes: A01N 57/16 containing heterocyclic radicals
• CPC Class Codes: A01N 63/40 Viruses, e.g. bacteriophages
• CPC Class Codes: A01N 63/50 Isolated enzymes; Isolated proteins peptides A01N37/46
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8218 Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
• CPC Class Codes: C12N 2310/12 catalytic nucleic acids, e.g. ribozymes
• CPC Class Codes: C12N 2310/121 Hammerhead
• CPC Class Codes: C12N 2310/124 based on group I or II introns

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 target RNA, a region of defined sequence and a base paired stem region.

Claims

1. A compound having the formula:

##STR10## wherein each X represents a ribonucleotide which may be the same or different;

wherein each of (X)_n-1 and (X)_n'"'"' represents an oligoribonucleotide having a predetermined sequence which is (a) capable of hybridizing with an RNA target sequence to be cleaved and (b) does not naturally occur covalently bound to the sequences C--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-1 and n'"'"' represents an integer which defines the number of ribonucleotides in the oligonucleotide with the proviso that the sum of n+n'"'"' is greater than or equal to 14;

wherein each * represents base pairing between the ribonucleotides 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; and

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 2 if (X)_b is present.

2. A compound of claim 1, wherein each of n and n'"'"' is greater than or equal to 6.

3. A compound having the formula:

##STR11## wherein Q represents a compound of claim 1;

wherein each Y represents a ribonucleotide which may be the same or different;

wherein each of r and s represents an integer which may be greater than or equal to 0; and

wherein z represents an integer greater than or equal to 1.

4. A composition which comprises a compound of any of claims 1, 2 or 3 in association with a pharmaceutically, veterinarially, or agriculturally acceptable carrier or excipient.

5. 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 any of claims 1, 2 or 3.

6. A transfer vector according to claim 5, which is a bacterial plasmid or phage DNA.

7. A prokaryotic or eukaryotic cell containing a nucleotide sequence which on transcription gives rise to a compound of any of claim 1, 2 or 3.