Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2005-03-01
2005-03-01
Ponnaluri, Padmashri (Department: 1639)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S029000, C435S320100, C435S481000, C435S091500, C435S091500, C435S091500, C435S091500, C435S069100, C530S350000, C536S023500, C536S023100, C536S025320, C536S023400, C536S018700
Reexamination Certificate
active
06861220
ABSTRACT:
A method for screening molecule which have a synthetic lethal property when in combination with a gene of interest carrying a non-lethal mutation, said method comprising the steps of: transfecting a first reporter gene into mammalian cells having a genome comprising a gene of interest which carries a non-lethal mutation, or a genome which is null of said gene of interest; selecting clones stably expressing said fist reporter gene; introducing into said cells a survival plasmid comprising a functioning copy of said gene of interest, a second reporter gene, selectable marker, an origin of DNA replication and a nuclear antigen gene essential for replication of the plasmid within said cells, wherein said survivsal plasmid is autonomously replicating and spontaneously lost from said cells; growing said cells in the presence of a selection compound which selects for said selectable marker; selecting cell clones stably expressing said second reporter gene and said functioning copy of said gene of interest; removing selection for the selectable marker, and adding molecules destined for screening of their ability to impose selective pressure enforcing retention of the unstable survival plasmid, determining survival plasmid retention in cells, thus identifying a molecule having a synthetic lethal property when in combination with non lethal mutated gene of interest.
REFERENCES:
patent: 4686186 (1987-08-01), Sugden
patent: 5168062 (1992-12-01), Stinski
patent: 5625048 (1997-04-01), Tsien et al.
Deiss et al., Science, vol. 252, pp. 117-120, Apr. 1991.*
Wade-Martins et al., Nucleic acid Research, vol. 27, No. 7, pp. 1674-1682, 1999.*
Simons A, Dafni N., Dotan I., Oron Y., Canaani D., “Establishment of a Chemical Synthetic Lethality Screen in Cultured Human Cells,” Genome Research p. 266 www.genome.org/cgi/doi/10.1101/gr.154201.
Yates, J., Warren N. Sugden B., “Stable replications of plasmids derived from Epstein-Barr virus in various mammalian cells” Nature vol. 313 2B Feb. 1985.
Zhu Z., Geertruida M., Veldman., Cowie A., Carr A., Schaffhausen B., Kamen R., “Construction and Functional Characterization of Polyomavirus genomes that separately encode the three early proteins” Journal of Virology, Jul. 1984 p. 170-180 vol. 51 1984.
Gonzalez J.E., Negulescu P.A., “Intracellular detection assays for high throughput screening” Current opinion in Biotechnology 1998, 9:624-631.
Canaani Dan
Dafni Naomi
Dotan Iris
Simons Arnold
Cohen Mark S.
Ponnaluri Padmashri
Ramot University Authority for Applied Research & Industria
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