Claims for Patent: 6,713,257

Summary for Patent: 6,713,257

Title:	Gene discovery using microarrays
Abstract:	The invention relates to methods and systems (e.g., computer systems and computer program products) for identifying and characterizing genes using microarrays. In particular, the invention provides for improved, robust methods for detecting genes through the use of microarrays to analyze the expression state of the genome. Genes which are expressed can be mapped to their respective positions in the genome, and the structure of such genes can be determined.
Inventor(s):	Shoemaker; Daniel D. (Bothell, WA), Scherer; Stewart (Moraga, CA), Altschuler; Steven J. (Redmond, WA), Wu; Lani F. (Redmond, WA), Armour; Christopher D. (Kirkland, WA)
Assignee:	Rosetta Inpharmatics LLC (Kirkland, WA)
Application Number:	09/781,814
Patent Claims:	1. A method of identifying the location of exons within the genome of a species of organism comprising: (a) contacting a sample comprising RNAs or nucleic acids derived therefrom from one or more cells of said species of organism with an array, said array comprising a positionally-addressable ordered array of polynucleotide probes bound to a solid support, said polynucleotide probes comprising a first plurality of at least 100 polynucleotide probes of different, predetermined nucleotide sequences, each said different nucleotide sequence comprising a sequence complementary and hybridizable to a different genomic sequence of the same species of organism, said respective genomic sequences for the probes being found at sequential predetermined sites in said genome of said species of organism, said contacting being under conditions conducive to hybridization between said RNAs or nucleic acids derived therefrom and said probes; (b) identifying the one or more probes to which hybridization of one or more of said RNAs or nucleic acids derived therefrom occurs; and (c) identifying said genomic sequences for each said identified probe as the location of an exon within the genome of said species of organism. 2. The method of claim 1, wherein step (a) is repeated with RNAs or nucleic acids derived therefrom from a plurality of different cells of said species of organism. 3. The method of claim 1, wherein said array has in the range of 150 to 1,000 different polynucleotide probes per 1 cm.sup.2. 4. The method of claim 1, wherein said array has in the range of 1,000 to 10,000 different polynucleotide probes per 1 cm.sup.2. 5. The method of claim 1, wherein said array has in the range of 10,000 to 50,000 different polynucleotide probes per 1 cm.sup.2. 6. The method of claim 1, wherein said array has greater than 50,000 different polynucleotide probes per 1 cm.sup.2. 7. The method of claim 1, wherein the nucleotide sequences of the probes consist of up to 1000 nucleotides. 8. The method of claim 1, wherein the probes are 10-200 nucleotides in length. 9. The method of claim 1, wherein the probes are 80-120 nucleotides in length. 10. The method of claim 1, wherein the probes are 40-80 nucleotides in length. 11. The method of claim 1, wherein the nucleotide sequences of the probes consist of 60 nucleotides. 12. The method of claim 1, wherein said genomic sequences for different probes are overlapping in said genome. 13. The method of claim 1, wherein said genomic sequences for different probes are overlapping in said genome from 10-50% of the length of each said different probe. 14. The method of claim 1, wherein said genomic sequences for different probes are adjacent in said genome. 15. The method of claim 1, wherein said genomic sequence for each probe is spaced apart from that for other probes in said genome by less than 200 bp. 16. The method as in one of claims 7-11, wherein said genomic sequences for different probes are overlapping in said genome. 17. The method as in one of claims 7-11, wherein said genomic sequences for different probes are overlapping in said genome from 10-50% of the length of each said different probe. 18. The method as in one of claims 7-11, wherein said genomic sequences for different probes are adjacent in said genome. 19. The method as in one of claims 7-11, wherein said genomic sequence for each probe is spaced apart from that for other probes in said genome by less than 200 bp. 20. The method of claim 1, wherein said organism is a eukaryote. 21. The method of claim 1, wherein said organism is a human. 22. The method of claim 1, wherein said organism is a plant. 23. The method of claim 1, wherein said organism is a mammal. 24. The method of claim 1, wherein said first plurality of polynucleotide probes is at least 1,000 probes. 25. The method of claim 1, wherein said first plurality of polynucleotide probes is at least 10,000 probes. 26. The method of claim 1, wherein said first plurality of polynucleotide probes is in the range of 1,000 to 50,000 probes. 27. The method of claim 1, wherein two or more of said polynucleotide probes are complementary and hybridizable to intron sequences of at least 10 different genes. 28. The method of claim 1, wherein the distance between 5' ends of said sequential sites is always less than 500 bp, and wherein the genomic sequences for said first plurality of probes span a genomic region of at least 25,000 bp. 29. The method of claim 1, wherein two or more of said polynucleotide probes are complementary and hybridizable to sequences contained entirely within an intron, and herein said ordered array does not comprise a second plurality of polynucleotide probes hat do not comprise a sequence complementary and hybridizable to said genome of said species of organism, said second plurality being of equal or greater number than said first plurality. 30. The method of claim 1, wherein two or more of said polynucleotide probes are complementary and hybridizable to intron sequences of at least 10 different genes, and wherein said ordered array does not comprise a second plurality of polynucleotide probes that do not comprise a sequence complementary and hybridizable to said genome of said species of organism, said second plurality being of equal or greater number than said first plurality. 31. The method of claim 1, wherein: (a) said polynucleotide probes further comprise a second plurality of polynucleotide probes comprising a sequence complementary and hybridizable to said first plurality; and (b) said identifying step comprises using a hybridization signal generated in said contacting step from said second plurality to filter a hybridization signal generated in said contacting step from said first plurality. 32. The method of claim 1, wherein: (a) said sample comprises RNAs or nucleic acids derived therefrom from (i) a first cell or cells of a first tissue type or subject to a first condition, and (ii) a second cell or cells of a second tissue type different from said first tissue type or subject to a second condition different from said first condition; and (b) said identifying step comprises comparing a hybridization signal generated in said contacting step from said first cell or cells to a hybridization signal generated in said contacting step from said second cell or cells. 33. The method of claim 1, wherein said plurality of probes is tiled across a sequence predicted to contain, or known to contain, exons. 34. The method of claim 1, wherein said plurality of probes includes known expressed sequence tags (ESTs) or predicted exons. 35. The method of claim 1, wherein each of said plurality of probes is complementary and hybridizable to a predicted or known exon. 36. The method of claim 1, further comprising a sample comprising a population of cellular RNA or nucleic acid derived therefrom on the surface of said solid support such that said sample is in contact with said polynucleotide probes, under conditions conducive to hybridization between said population and said polynucleotide probes. 37. The method of claim 36 wherein said population is labeled. 38. The method of claim 36 wherein said population comprises total cellular mRNA or nucleic acid derived therefrom. 39. The method of claim 36 wherein said population comprises nucleic acids of at least 10,000 different sequences. 40. A computer system for identifying the location of exons within the genome of a species of organism, said computer system comprising: one or more processor units; and one or more memory units connected to said one or more processor units, said one or more memory units containing one or more programs which cause said one or more processor units to execute steps of: (a) receiving a first data structure comprising a first plurality of measured hybridization signals from an array comprising a positionally-addressable ordered array of polynucleotide probes bound to a solid support, said polynucleotide probes comprising a second plurality of at least 100 polynucleotide probes of different, predetermined nucleotide sequences, each said different nucleotide sequence comprising a sequence complementary and hybridizable to a different genomic sequence of the same species of organism, said respective genomic sequences for the probes being found at sequential predetermined sites in said genome of said species of organism, said array contacting a sample comprising RNAs or nucleic acids derived therefrom from one or more cells of said species of organism with said array, said contacting being under conditions conducive to hybridization between said RNAs or nucleic acids derived therefrom and said probes; (b) receiving a second data structure comprising the nucleotide sequence of said genome of said organism; (c) receiving a third data structure comprising the nucleotide sequence of said second plurality of polynucleotide probes, said third data structure identifying the positional location of each said probe on said array; (d) identifying the one or more probes to which hybridization of one or more of said RNAs or nucleic acids derived therefrom occurs; (e) identifying said genomic sequences for each said identified probe as the location of an exon within the genome of said species of organism; and (f) outputting the locations of said exons with respect to the nucleotide sequence of said genome of said organism. 41. A computer program product for identifying the location of exons within the genome of a species of organism, the computer program product for use in conjunction with a computer having a memory and a processor, the computer program product comprising a computer readable storage medium having a computer program mechanism encoded thereon, wherein said computer program mechanism can be loaded into the one or more memory units of a computer and cause the one or more processor units of the computer to execute the steps of: (a) receiving a first data structure comprising a first plurality of measured hybridization signals from an array comprising a positionally-addressable ordered array of polynucleotide probes bound to a solid support, said polynucleotide probes comprising a second plurality of at least 100 polynucleotide probes of different, predetermined nucleotide sequences, each said different nucleotide sequence comprising a sequence complementary and hybridizable to a different genomic sequence of the same species of organism, said respective genomic sequences for the probes being found at sequential predetermined sites in said genome of said species of organism, said array contacting a sample comprising RNAs or nucleic acids derived therefrom from one or more cells of said species of organism with said array, said contacting being under conditions conducive to hybridization between said RNAs or nucleic acids derived therefrom and said probes; (b) receiving a second data structure comprising the nucleotide sequence of said genome of said organism; (c) receiving a third data structure comprising the nucleotide sequence of said second plurality of polynucleotide probes, said third data structure identifying the positional location of each said probe on said array; (d) identifying the one or more probes to which hybridization of one or more of said RNAs or nucleic acids derived therefrom occurs; (e) identifying said genomic sequences for each said identified probe as the location of an exon within the genome of said species of organism; and (f) outputting the locations of exon boundaries with respect to the nucleotide sequence of said genome of said organism.

Details for Patent 6,713,257

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2020-08-25
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2020-08-25
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2020-08-25