Hybrid adeno-retroviral vector for the transfection of cells

Abstract

An adenovirus, including adenoviral capsid proteins, and a replication-defective adenoviral vector that includes a 5′ retroviral LTR nucleic acid sequence, a 3′ retroviral LTR nucleic acid sequence, a nucleic acid sequence encoding a portion of a retroviral envelope protein adjacent to either the 5′ LTR or the 3′ LTR nucleic acid sequence, a retroviral packaging sequence and a nucleic acid sequence encoding a transgene located between the 5′ LTR and the 3′ LTR is provided. Host cells infected with this adenovirus are also provided. An adenoviral vector is provided that includes an adenoviral polynucleotide sequence comprising a nucleic acid encoding a transgene, a retroviral packaging signal, a 5′ and a 3′ retroviral LTR, and a portion of a retroviral envelope polypeptide, wherein the adenoviral polynucleotide sequence does not encode one or more of E1, E3 or E4. A method for transforming a cell is also provided using a virus or a vector of the invention, as is a method for introducing a transgene into a cell that is not able to produce viral particles with a single viral vector. A method is also provided for preventing or treating disorder in a subject using the adenoviral vectors of the invention. A pharmaceutical composition is also provided that includes an adenoviral vector of the invention and a pharmaceutically acceptable carrier.

Description

CROSS REFERENCE TO RELATED APPLICATION[0001]This is a continuation of U.S. patent application Ser. No. 11 / 255,059, filed Oct. 19, 2005, which is continuation of U.S. patent application Ser. No. 10 / 182,644, filed on Jul. 30, 2002, now U.S. Pat. No. 7,052,904, which is the §371 U.S. National Stage of International Application No. PCT / US01 / 03026, filed Jan. 30, 2001, which was published in English under PCT Article 21(2), and claims the benefit of U.S. Application No. 60 / 179,327, filed Jan. 31, 2000. The entire disclosure of each of these applications is hereby expressly incorporated by reference.FIELD OF THE INVENTION[0002]This invention relates to the field of viral vectors and the transduction of cells, more specifically to hybrid adenoviral vectors and their use in the transduction of cells in vitro or in vivo.BACKGROUND[0003]The transfer of genes into cells provides a means to determine gene function and treat diseases of genetic basis. In addition, gene transfer provides the basi...

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Benefits of technology

[0021]A method is also provided for preventing or treating a disorder in a subject, such as a genetic disorder, for example a disorder caused by a failure to adequately express a gene product. The method includes introducing into a cell of the subject a therapeutically effective amount of an adenoviral vector of the invention, wherein the cell is unable to produce viral particles. The introduction of the hybrid adenoviral vector results in the stable genetic transfection of the cell and expression of the transgene, such that a symptom of the disorder is alleviated or the disorder is prevented.

Problems solved by technology

However, this method was inefficient for the introduction of nucleic acid into many types of cells.

However, neither of these methods allow the transfer of nucleic acids into a cell with a high efficiency, nor do they allow for persistent gene expression.

Although adenoviral vectors, including gutless vectors, can infect both dividing and non-dividing cells, they generally do not stably integrate into the cellular genome.

However, retroviruses can transform only cells that are dividing; retroviral vectors cannot be used to introduce nucleic acid into non-dividing cells.

However, since the recombinant vector produced is a retrovirus, cell division is still required for virus infection and integration.

This latter fact is a significant drawback to target cells that are terminally differentiated and non-dividing.

In addition, each of these systems requires the use of more than one virus.

However, the efficiency of gene transfer and expression in vivo is still relatively low (Donehower, et al., Proc. Natl. Acad. Sci.

The gene delivery system is frequently the limiting factor for successful gene therapy.

Unfortunately, none of the vector systems presently in use meet all of these requirements.

Although adenoviruses can be produced at very high titers and may infect cells with high efficiency, they integrate into the cell genome only at very low frequency, which results in unstable gene expression.

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