# US 5,707,840 A · Multifunctional RNA having self-processing activity, the preparation thereof and the use thereof

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- Number: US 5,707,840 A
- Document ID: US5707840A
- Status: Granted
- Kind: A
- Application number: 08/313,608
- Filed: 1994-09-29
- Granted: 1998-01-13
- Earliest claimed: 1989-10-06
- Assignees: Hoechst Aktiengesellschaft
- Inventors: Hubert Mullner; Eugen Uhlmann; Peter Eckes; Rudolf Schneider; Bernadus Uijtewaal
- Web page: https://patentgrep.com/patent/US5707840A

## CPC

- 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)}
- C12N 15/1131: {against viruses}
- C12N 15/8283: {for virus resistance}
- C12N 2310/111: spanning the whole gene, or a large part of it
- C12N 2310/12: catalytic nucleic acids, e.g. ribozymes
- C12N 2310/121: Hammerhead
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A multifunctional RNA having self-processing activity, the preparation thereof and the use thereof. Host cells can be transformed so that they express ribozyme RNA and antisense RNA which are connected with each other via a spacer. The RNA molecules can, for example, be complementary to a certain viral RNA. Plants which have been transformed with genes coding for RNA of this type show a significantly improved resistance to viruses.

## Claims

### Claim 1

An isolated DNA fragment which encodes a pattern which comprises:
5'-hammerhead ribozyme RNA-(spacer-antisense RNA).sub.n -spacer-hammerhead ribozyme RNA-3', wherein n is a number from 0 to 10, and the spacer is a chain of 20 to 25 nucleotides which contains a ribozyme cleavage site in each case; and wherein said DNA fragment encodes a multifunctional RNA with self-processing activity.

### Claim 2 (depends on 1)

The isolated DNA fragment as claimed in claim 1, wherein the encoded antisense RNA is complementary to a vital RNA.

### Claim 3 (depends on 1)

The isolated DNA fragment as claimed in claim 1, wherein the encoded ribozyme mRNA has at least 6 consecutive nucleotides at each of the 5' and 3' ends that are complementary to a viral RNA.

### Claim 4

The isolated DNA fragment which comprises the DNA SEQUENCE ID NO. 1.

### Claim 5 (depends on 1)

A multifunctional RNA encoded by the isolated DNA fragment as claimed in claim 1.

### Claim 6 (depends on 1)

A plant host cell containing the DNA fragment as claimed in claim 1.

### Claim 7 (depends on 5)

A plant host cell containing an RNA as claimed in claim 5.

### Claim 8 (depends on 6)

The plant host cell of claim 6, wherein said plant host cell is a dicotyledonous plant host cell.

### Claim 9 (depends on 7)

The plant host cell of claim 7, wherein said plant host cell is a dicotyledonous plant host cell.

### Claim 10 (depends on 6)

A multifunctional RNA as claimed in claim 6, wherein the RNA is encoded by the isolated DNA fragment comprising the sequence in SEQUENCE ID NO. 1.