Methods and compositions for expressing negative-sense viral rna in canine cells - Patents

Methods and compositions for expressing negative-sense viral rna in canine cells
Agent: Johnathan Klein-evans - Gaithersburg, MD, US
Inventors: Gregory Duke, George Kemble, James Young, Zhaoti Wang
Applicaton #: 20060286591
Class: 435006000 (USPTO)
12/21/06
The present invention provides novel canine pol I regulatory nucleic acid sequences useful for the expression of nucleic acid sequences in canine cells such as MDCK cells. The invention further provides expression vectors and cells comprising such nucleic acids as well as methods of using such nucleic acids to make influenza viruses, including infectious influenza viruses.

FIELD OF THE INVENTION
[0002] In one aspect, the present invention provides an isolated nucleic acid comprising a canine RNA polymerase I regulatory sequence. In other aspects, the invention provides expression vectors and cells comprising such nucleic acids as well as methods of using such nucleic acids to make influenza viruses, including infectious influenza viruses.
SUMMARY
[0012] Disclosed herein are nucleic acids which comprise regulatory elements that can be used to express, for example, influenza genomic RNA in canine cells. Compositions such as isolated nucleic acids, vectors, and cells comprising the canine regulatory sequences of the invention, and methods of using the same are embodiments of the subject invention.
[0013] Accordingly, in certain aspects, isolated nucleic acids of the invention comprise a canine RNA polymerase I (pol I) regulatory sequence. In certain embodiments, the regulatory sequence comprises a promoter. In certain embodiments, the regulatory sequence comprises an enhancer. In certain embodiments, the regulatory sequence comprises both a promoter and an enhancer. In one embodiment, the regulatory sequence comprises nucleotides -250 to -1 (in relation to the first nucleotide transcribed from the promoter, also known as the +1 nucleotide) of the corresponding native promoter or a functional derivative thereof. In one embodiment, the regulatory sequence is operably linked to a viral DNA, e.g., a cloned viral cDNA. In one embodiment, the cloned viral cDNA encodes viral RNA of a negative or positive strand virus or the corresponding cRNA. In certain embodiments, the cloned viral cDNA encodes genomic viral RNA (or the corresponding cRNA) of an influenza virus.
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