Cornell Research Foundation Inc.

Innovir Laboratories, Inc.

Hugh Robertson

Hazel Szeto

Kesheng Zhao

Alex V Birk

Allan R Goldberg

Shaji T George

Thomas J Kindt

Timothy W Nilsen

The invention relates to carrier complexes and methods for delivering molecules to cells. The carrier complexes comprises a molecule and an aromatic cationic peptide in accordance with the invention. In one embodiment, the method for delivering a molecule to a cell comprises contacting the cell with a carrier complex. In another embodiment, the method for delivering a molecule to a cell comprises contacting the cell with a molecule and an aromatic cationic peptide.

Method and carrier complexes for delivering molecules to cells

Vectors and a method for the identification of affector RNA molecules, such as ribozymes, external guide sequences, anti-sense RNA, and triple helix-forming RNA, that inhibit expression of target RNA molecules are disclosed. The method identifies functional affector RNA molecules by screening or selecting for those RNA molecules that inhibit expression of a fusion transcript, which includes the sequence of an RNA molecule of interest, from a library of potential affector RNA molecules. The vectors include a reporter gene encoding the fusion transcript including the RNA molecule of interest and RNA encoding the reporter protein. The vectors also include a second reporter gene encoding a second reporter protein. Expression of the second reporter protein can be used both to detect transformation or transfection of the vector into cells and as a control for effects on the expression of the first reporter protein that are not due to inhibition of expression of the RNA molecule of interest. The vector also encodes an affector RNA molecule targeted to the RNA of interest. A key advantage of the disclosed method and vectors is the assessment of inhibition of expression of an RNA of interest in an in vivo setting which will be the same or similar to the setting where identified affector molecules will be used. Another advantage of the disclosed method is that all, or a substantial number of the accessible sites in the RNA of interest can be determined in one assay. Also disclosed are affector oligomers based on affector RNA molecules identified as inhibiting the expression of an RNA of interest. The disclosed method also allows direct comparison of the inhibitory activities of different affector RNA molecules directed to different target sites.

Random intracellular method for obtaining optimally active nucleic acid

Hepatitis delta is used as a vector for inhibition of viral infection and to express proteins in vivo in a cell-specific manner. The scope of delta's use as a vector is broadened in the present invention in several important ways. For example, a delta RNA genome capable of self-replication is enlarged to carry additional information, either coding for messenger RNA for a protein, or for a targeted ribozyme, which can be delivered to liver cells using delta's normally infectious properties, or to other cell types using chimeric delta viral agents carrying altered surface proteins. In another embodiment, the delta vector is made self-limiting, so that its role in delivering targeted information is separated from its viral property of unlimited infectious replication. Targeting is achieved through the use of sequences flanking the delta sequences that have affinity for sites on RNA to be cleaved.

Ribozyme compositions and expression vectors

Hepatitis delta virus compositions and expression vectors

Ribozymes, sequences cleaving RNA, derived from sequences present in the hepatitis delta virus, have been engineered for greater specificity without increasing size. The specific ribozyme sequences are useful as reagents for cleaving RNA for experimental studies as well as antiviral therapies. Examples demonstrating the targeting of these sequences against HIV and Crohn's disease are described in detail. The sequences are also useful as diagnostics for the detection of hepatitis delta virus in tissue and fluid samples, as in blood banking, as well as in isolation and characterization of new viroids having ribozyme activity, using an RNA-specific hybridization method. Based on analysis of the two domain structure of the hepatitis delta virus, it is possible to construct a vector for expression of non-hepatitis delta virus proteins in mammalian cells.

Ribozyme compositions and methods for use

Therapeutic ribozyme compositions and expression vectors

Growing community of inventors

New York, NY, United States of America

Hugh Robertson