EP0387775B1
Expired / Lapsed
EP
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Genetic construct for inhibiting RNA function
Assignee: Boehringer Ingelheim International GmbH
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Filing: 1990-03-13
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Granted: 1996-07-31
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Expires: 2010-03-13
Abstract
Abstract The invention relates to a genetic unit, which is optionally in multicopy form, for the inhibition of RNA. The unit contains the transcription units necessary for transcription by polymerase III, and a DNA which codes for inhibiting RNA and which is arranged within the unit in such a way that the transcribed RNA is part of the polymerase III transcript. These units can be used to achieve increased stability of the inhibiting RNA, which can be in the form of ribozymes or antisense RNAs, while there is no reduction in the activity. The invention furthermore relates to a process for introducing the genetic units into the cell, to the use of these units and to pharmaceutical formulations containing them.
Inventors
- 👤 Hartmut Dr. Beug
- 👤 Max L. Prof.-Dr. Birnstiel
- 👤 Matthew Dr. Cotten
- 👤 Ernst Dr. Wagner
- 👤 Harald Dipl.-Ing. Kandolf
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 05:57
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; A; A61; A61K; A61K38/00; C07; C07K; C07K2319/00; C12N2310/00; C12N2310/10; C12N2310/11; C12N2310/111; C12N2310/12; C12N2310/121
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
87 words
DNA molecule, optionally occurring in multiple copies, containing sections of a gene transcribed by polymerase III and a DNA sequence coding for an inhibiting RNA molecule, characterized in that it contains the transcription units of a tRNA gene necessary for transcription by polymerase III, including the sequences which determine the secondary structure of the tRNA, and that the DNA sequence coding for an inhibiting RNA molecule is arranged inside the DNA molecule in such a way that the inhibiting RNA molecule is a part of the transcript.
Claim 2
Dependent Claim
16 words
DNA molecule according to claim 1, characterized in that the inhibiting RNA molecule is a ribozyme.
Claim 3
Dependent Claim
18 words
DNA molecule according to claim 2, characterized in that the ribozyme is a ribozyme of the hammerhead type.
Claim 4
Dependent Claim
17 words
DNA molecule according to claim 1, characterized in that the inhibiting RNA molecule is an antisense RNA.
Claim 5
Dependent Claim
18 words
DNA molecule according to claim 1, characterized in that it contains the transcription units of an initiation-met tDNA.
Claim 6
Dependent Claim
18 words
DNA molecule according to claim 1, characterized in that it contains the transcription units of a tyr tDNA.
Claim 7
Dependent Claim
38 words
DNA molecule according to one of claims 1 to 6, characterized in that it contains the DNA sequence coding for the inhibiting RNA molecule as an insert between the A block and B block of the tRNA gene.
Claim 8
Dependent Claim
37 words
DNA molecule according to claims 5 or 7, characterized in that it contains the DNA sequence coding for the inhibiting RNA molecule as an insert in the natural Apa1-restriction cutting site between the A and B blocks.
Claim 9
Dependent Claim
26 words
DNA molecule according to claim 6, characterized in that it contains the DNA sequence coding for the inhibiting RNA molecule as an insert in the intron.
Claim 10
Dependent Claim
36 words
DNA molecule according to one of the preceding claims, characterized in that it contains an oligonucleotide sequence such that the anticodon stem region of the tRNA transcript is longer than that of the wild-type tRNA transcript.
Claim 11
Dependent Claim
32 words
DNA molecule according to claim 9 or 10, characterized in that it contains the DNA sequence coding for the inhibiting RNA molecule as an insert in an artificially introduced restriction cutting site.
Claim 12
Dependent Claim
36 words
DNA molecule according to claims 9 to 11, characterized in that the intron is modified so that the secondary structure of the precursor tRNA is stabilized, whilst the structures which are critical for splicing are maintained.
Claim 13
Dependent Claim
22 words
DNA molecule according to one of claims 1 to 12, characterized in that the inhibiting RNA molecule is directed against viral RNA.
Claim 14
Dependent Claim
33 words
DNA molecule according to one of claims 1 to 12, characterized in that the inhibiting RNA molecule is directed against oncogenes or other key genes which control the growth and/or differentiation of cells.
Claim 15
Dependent Claim
38 words
DNA molecule according to one of claims 1 to 14, characterized in that it occurs as a multiple copy, whilst identical or different sub-units containing DNA coding for an inhibiting RNA molecule are present as complete transcription units.
Claim 16
Dependent Claim
47 words
Process for introducing DNA molecules according to one of claims 1 to 15 into higher eukaryotic cells, characterized in that the DNA molecules are modified so that they are absorbed into the cell by receptor-mediated endocytosis and the cells are brought into contact with the DNA molecules.
Claim 17
Dependent Claim
27 words
Process according to claim 16, characterized in that the DNA molecules are complexed with a transferrinpolycation conjugate and the cells are brought into contact with this complex.
Claim 18
Dependent Claim
19 words
Use of the DNA molecules defined in one of claims 1 to 15 for inhibiting RNA in vivo .
Claim 19
Dependent Claim
19 words
Use according to claim 18 for inhibiting oncogenes or other key genes which control growth and/or differentiation of cells.
Claim 20
Dependent Claim
9 words
Use according to claim 18 for inhibiting pathogenic viruses.
Claim 21
Dependent Claim
17 words
Use according to claim 18 for producing transgenic animals in which the DNA molecule is constitutively transcribed.
Claim 22
Dependent Claim
19 words
Use according to claim 18 for the preparation of transgenic plants in which the DNA molecule is constitutively expressed.
Claim 23
Dependent Claim
19 words
Pharmaceutical preparation containing as active component one or more DNA molecules according to one of claims 1 to 15.
Claim 24
Dependent Claim
10 words
Pharmaceutical preparation according to claim 23 for inhibiting viral RNA.
Claim 25
Dependent Claim
20 words
Pharmaceutical preparation according to claim 23 for inhibiting oncogenes or other key genes which control the growth rate of cells.
Claim 26
Dependent Claim
14 words
Pharmaceutical preparation according to claim 23 for inhibiting the synthesis of undesirable gene products.
Claim 27
Dependent Claim
25 words
Pharmaceutical preparation according to one of claims 23 to 26, characterized in that it additionally contains a transferrin-polycation conjugate capable of complexing the DNA molecule.
Claim 28
Dependent Claim
14 words
Pharmaceutical preparation according to claim 27, characterized in that it contains a transferrinpolylysine conjugate.
Claim 29
Dependent Claim
14 words
Pharmaceutical preparation according to claim 27, characterized in that it contains a transferrinprotamine conjugate.
References Cited (Backward Citations)
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