Stable and long-lasting siRNA expression vectors and the...

Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives

Reexamination Certificate

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C435S320100, C435S325000

Reexamination Certificate

active

07973155

ABSTRACT:
The invention relates to a siRNA expression vector that can inhibit or eliminate the expression of a target gene in a mammalian cell, said vector comprising: a bacterial cassette containing a bacterial origin of replication and a bacterial selection marker M1; a eucaryotic cell selection cassette comprising a marker M2 for selecting eucaryotic cells under the control of an appropriate promoter; a cassette EBV comprising at least one fragment of the antigen EBNA-1, at least one fragment FR, and at least one fragment of the region DYAD; and a siRNA transcription cassette comprising at least one region coding for a siRNA corresponding to the target gene to be inhibited or eliminated, under the control of elements for regulating transcription in mammalian cells, said regulating elements including at least one promoter capable of transcribing a siRNA in mammalian cells, and a transcription terminator. The invention also relates to the applications of one such expression vector.

REFERENCES:
patent: 1 484 393 (2004-12-01), None
Biard, Denis S. F. et al.,“ Development of New EBV-Based for Stable Expression of Small Interfering RNa to Mimick Human Syndromes: Application to NNER Gene Silencing”, Mol. Cancer Rec., vol. 3, No. 9. pp. 519-529, 2005. XP002349462.
Miyagishi, M. et al.,“ Expressions of siRNA from a pol III Promoter in Mammalian Cells”, Perspectives in Gene Expression, pp. 361-375, 2003. XP001246949.
Brummelkamp, T. R. et al.,“A System for Stable Expression of Short Interfering RNAs in Mammalian Cells”, Science, vol. 296, No. 5567, pp. 550-553, 2002. XP002225638.
Database Google “Invitrogen Life Technologies” HTTP://WWW. INVITROGEN.COM/CONTENT/SFS/VECTORS/PCDNA3.PDF, PDF Data Sheet, 2006. XP002386946.
Biard, D. S. F. et al.,“Regulation of theEscherichia ColiLac Operon Expressed in Human Cells”, Biochimica et Biophisica Acta, vol. 1130, No. 1, pp. 68-74, 1992. XP009055483.
Biard, D. S. F. et al.,“Establishment of a Human Cell Line for the Detection of Demethylating Agents”, Experimental Cell Research, vol. 200, No. 2, pp. 263-271, 1992. XP 009055400.
Chittenden, T. et al.,“Functional Limits of oriP, the Epstein-Barr Virus Plasmid Origin of Replication”, Joumal of Virology, vol. 63, No. 7, pp. 3016-3025, 1989. XP 002914212.

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