1. A method of facilitating zero sign-on access to media services comprising:
providing a trust credential to a client application operating on a user device, the trust credential specifying media service permissions for the user device; and
allowing the user device zero sign-on access to media services in accordance with the trust credential specified media service permissions.
2. The method of claim 1 further comprising:
configuring the trust credential to facilitate access to a first tier of media service permissions in the event an access point of the user device is trusted; and
configuring the trust credential to facilitate access to a second tier of media service permissions in the event the access point of the user device is untrusted.
3. The method of claim 2 further comprising the first tier of media service permissions corresponding with subscriber specific services and the second tier of media service permissions corresponding with non-subscriber specific services.
4. The method of claim 2 further comprising:
determining the access point to be trusted in the event an IP address of the user device provided through the access point is within a trusted domain of IP addresses; and
determining the access point to be untrusted in the event the IP address of the user device provided through the access point is within an untrusted domain of IP addresses.
5. The method of claim 1 further comprising relating the media service permissions to a subscriber associated with the access point.
6. The method of claim 5 further comprising the media service permissions of the subscriber associated with the access point being different than a subscriber associated with the user device.
7. The method of claim 1 further comprising relating the media service permissions to a subscriber associated with the user device.
8. The method of claim 1 further comprising configuring the trust credential as a cookie to be stored in a computer-readable medium of the user device.
9. The method of claim 8 further comprising requiring a refresh of the cookie no more than once every period in order to continue zero sign-on access to media services, the period being less than six months.
10. The method of claim 1 further comprising configuring the trust credential as a certificate to be stored in a computer-readable medium of the user device.
11. The method of claim 10 further comprising requiring a refresh of the certificate no more than once every period in order to continue zero sign-on access to media services, the period being greater than six months.
12. A method of facilitating zero sign-on access to media services comprising:
determining whether an access point through which a user device desires to access the media services is one of trusted and untrusted;
in the event the access point is trusted, allowing zero sign-on access to the media services; and
in the event the access point is untrusted, determining whether a trust credential stored on the user device is one of valid and invalid;
i. in the event the trust credential is valid, allowing zero sign-on access the media services; and
ii. in the event the trust credential is invalid, requiring sign-on access to the media services.
13. The method of claim 12 further comprising limiting the zero sign-on access to media permissions specified in the trust credential.
14. The method of claim 13 further comprising relating the media permissions to a subscriber associated with the access point.
15. The method of claim 13 further comprising relating the media permissions to a subscriber associated with the user device.
16. The method of claim 12 further comprising:
determining the access point to be trusted in the event an IP address of the user device provided through the access point is within a trusted domain of IP addresses; and
determining the access point to be untrusted in the event the IP address of the user device provided through the access point is within an untrusted domain of IP addresses.
17. A method of facilitating zero sign-on access of a user device to media services comprising:
determining whether a trust credential stored on a user device is one of a first type and a second type;
in the event the trust credential is the first type, allowing zero sign-on access to a first tier of the media services; and
in the event the trust credential is the second type, allowing zero sign-on access to a second tier of the media services.
18. The method of claim 17 further comprising:
determining the trust credential is the first type in the event a username and password associated with the user device is unverified;
determining the trust credential is the second type in the event the username and password associated with the user device is verified.
19. The method of claim 17 further comprising:
relating the first tier of media services to an access point of the user device; and
relating the second tier of media services to the user device.
20. The method of claim 17 further comprising further limiting media services available under each of the first and second tiers in the event an access point of the user device is untrusted.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
We claim:
1. A pair of cells comprising:
a first cell; and
a second cell,
wherein the first cell and the second cell are isogenic but for:
a gene of interest and a gene encoding a fluorescent protein;
wherein the first cell comprises a gene that encodes a first fluorescent protein having a first absorption spectrum and a first emission spectrum;
wherein the second cell comprises a gene that encodes a second fluorescent protein having a second absorption spectrum and a second emission spectrum; and
wherein either:
the first and second absorption spectra are not identical; andor
the first and second emission spectra are not identical.
2. The pair of cells of claim 1 wherein the first and second absorption spectra are not identical and the first and second emission spectra are not identical.
3. The pair of cells of claim 1 wherein the cells are contained within the same undivided container.
4. The pair of cells of claim 1 wherein the first cell is homozygously wild-type for the gene of interest and wherein the second cell is homozygously mutant for the gene of interest.
5. The pair of cells of claim 1 wherein the gene of interest in the second cell is homozygously deleted.
6. The pair of cells of claim 1 wherein the first cell comprises two wild-type alleles of the gene of interest and wherein the second cell comprises a wild-type allele and a mutant allele of the gene of interest, wherein the mutant allele is dominant.
7. The pair of cells of claim 1 wherein the gene of interest is an oncogene and the first cell is homozygous for a mutant allele of the oncogene and wherein the second cell comprises a homozygous deletion of the mutant oncogene.
8. The pair of cells of claim 1 wherein the first cell expresses the gene of interest and wherein the second cell does not express the gene of interest.
9. The pair of cells of claim 1 wherein the first cell comprises a wild-type allele and a mutant allele of the gene of interest and the second cell is hemizygous for the wild-type allele of the gene of interest.
10. The pair of cells of claim 1 wherein the first cell expresses a protein encoded by the gene of interest and wherein the second cell does not express a protein encoded by the gene of interest.
11. The pair of cells of claim 1 wherein the first and second cells are mammalian cells.
12. The pair of cells of claim 1 wherein the first and second cells are human cells.
13. The pair of cells of claim 1 wherein the cells are cancer cells.
14. The pair of cells of claim 13 wherein the cancer cells are selected from the group consisting of colon tumor cells and breast tumor cells.
15. The pair of cells of claim 1 wherein the cells are HCT116 cells.
16. The pair of cells of claim 1 wherein the cells are DLD-1 cells.
17. The pair of cells of claim 1 wherein the first and second fluorescent proteins are selected from the group consisting of green fluorescent protein, red fluorescent protein, blue fluorescent protein, yellow fluorescent protein, and cyan fluorescent protein.
18. The pair of cells of claim 1 wherein the gene of interest is Ras and wherein the Ras genotype of the first cell is c-Ki-RasWTmutant and wherein the Ras genotype of the second cell is c-Ki RasWTnull.
19. A pair of cells comprising:
a first cell wherein the Ras genotype of the first cell is c-Ki-RasWTmutant and wherein the first cell comprises a first gene that encodes a first fluorescent protein having a first absorption spectrum and a first emisson spectrum; and
a second cell wherein the Ras genotype of the second cell is c-Ki-RasWTnull and wherein the second cell comprises a second gene that encodes a second fluorescent protein having a second absorption spectrum that is not identical to the first absorption spectrum and a second emission spectrum that is not identical to the first emission spectrum, wherein the first and second cells are isogenic but for the Ras gene and the gene encoding a fluorescent protein.
20. The pair of cells of claim 19 wherein the first fluorescent protein is blue fluorescent protein and the second fluorescent protein is yellow fluorescent protein.
21. A method of making a pair of cells, comprising the steps of:
genetically modifying a first cell to yield a second cell that is isogenic with the first cell but for a single gene of interest;
transfecting the first cell with a first gene that encodes a first fluorescent protein having a first absorption spectrum and a first emission spectrum; and
transfecting the second cell with a second gene that encodes a second fluorescent protein having a second absorption spectrum and a second emission spectrum, wherein either the first and second absorption spectra are not identical andor the first and second emission spectra are not identical.
22. The method of claim 21 wherein the first and second absorption spectra are not identical and wherein the first and second emission spectra are not identical.
23. The method of claim 21 wherein the first and second cells are mammalian cells.
24. The method of claim 21 wherein the first and second cells are human cells.
25. The method of claim 24 wherein the human cells are human cancer cells.
26. The method of claim 25 wherein the human cancer cells are selected from the group consisting of colon tumor cells and breast tumor cells.
27. The method of claim 21 wherein the first and second cells are HCT 116 cells.
28. The method of claim 21 wherein the first and second cells are DLD-1 cells.
29. The method of claim 21 wherein the Ras genotype of the first cell is c-Ki-RasWTmutant and wherein the Ras genotype of the second cell is c-Ki-RasWTnull.
30. A method of identifying a test compound as selectively affecting a gene of interest or its expression products or downstream genes or proteins in its pathway comprising the steps of:
culturing a first and second cell, wherein the first and second cells are isogenic but for a gene of interest and a gene encoding a fluorescent protein, wherein the first cell comprises a first gene that encodes a first fluorescent protein having a first absorption spectrum and a first emission spectrum, and wherein the second cell comprises a second gene that encodes a second fluorescent protein having a second absorption spectrum and a second emission spectrum, wherein either the first and second absorption spectra are not identical andor the first and second emission spectra are not identical;
contacting the first and second cells with a test compound; and
identifying the test compound as selectively affecting the gene of interest or its expression products or downstream genes or proteins in its pathway if the growth rate of the first cell is altered with respect to the growth rate of the second cell.
31. The method of claim 30 wherein the first and second cells are co-cultured.
32. The method of claim 30 wherein an equal number of the first and second cells are cultured.
33. The method of claim 30, wherein the first and second absorption spectra are not identical and wherein the first and second emission spectra are not identical.
34. The method of claim 30 wherein the fluorescent proteins are detected using fluorescence microscopy to assess growth rate.
35. The method of claim 30 wherein the fluorescent proteins are detected using high-throughput fluorescence spectroscopy to assess growth rate.
36. The method of claim 30 wherein the first cell is homozygously wild-type for the gene of interest and wherein the second cell is homozygously mutant for the gene of interest.
37. The method of claim 30 wherein the gene of interest in the second cell is homozygously deleted.
38. The method of claim 30 wherein the first cell comprises two wild-type alleles of the gene of interest and wherein the gene of interest in the second cell comprises a wild-type allele and a mutant allele of the gene of interest, wherein the mutant allele is dominant.
39. The method of claim 30 wherein the first cell is homozygous for a mutant oncogene and wherein the second cell comprises a homozygous deletion of the mutant oncogene.
40. The method of claim 30 wherein the first cell expresses the gene of interest and wherein the second cell does not express the gene of interest.
41. The method of claim 30 wherein the first cell comprises a wild-type allele and a mutant allele of the gene of interest and wherein the second cell is hemizygous for a wild-type allele of the gene of interest.
42. The method of claim 30 wherein the first cell expresses a mutant protein encoded by the gene of interest and wherein the second cell does not express a protein encoded by the gene of interest.
43. The method of claim 30 wherein the first and second cells are mammalian cells.
44. The method of claim 30 wherein the first and second cells are human cells.
45. The method of claim 30 wherein the cells are cancer cells.
46. The method of claim 45 wherein the cancer cells are selected from the group consisting of colon tumor cells and breast tumor cells.
47. The method of claim 30 wherein the first and second cells are HCT116 cells.
48. The method of claim 30 wherein the first and second cells are DLD-1 cells.
49. The method of claim 30 wherein the first and second fluorescent protein are selected from the group consisting of green fluorescent protein, red fluorescent protein, blue fluorescent protein, yellow fluorescent protein, and cyan fluorescent protein.
50. A method of identifying a test compound as selectively affecting a Ras gene, Ras protein, or downstream gene or protein in its pathway in cells comprising:
contacting a test compound with a co-culture of an essentially equal number of a first and a second cell that are isogenic but for their Ras genes and a gene encoding a fluorescent protein, wherein the Ras genotype of the first cell is c-Ki-RasWTmutant and wherein the first cell comprises a first gene encoding a first fluorescent protein having a first absorption spectrum and a first emisson spectrum and wherein the Ras genotype of the second cell is c-Ki-RasWTnull and wherein the second cell comprises a second gene encoding a second fluorescent protein having a second absorption spectrum that is not identical to the first absorption spectrum and a second emission spectrum that is not identical to the first emission spectrum; and
identifying the test compound as selectively affecting the Ras gene, Ras protein, or downstream gene or protein in the pathway if the growth rate of the first cell is altered with respect to the growth rate of the second cell.
51. The method of claim 50 wherein the first fluorescent protein is blue fluorescent protein and wherein the second fluorescent protein is yellow fluorescent protein.
52. A composition comprising at least 90% of a compound having a formula selected from the group consisting of:
4
53. A pharmaceutical composition comprising a compound with a formula selected from the group consisting of:
5
or pharmaceutically acceptable salts, solvates, or prodrugs thereof, and a pharmaceutically appropriate carrier.
54. A cytotoxic composition comprising a compound having a formula:
6
or a pharmaceutically acceptable salt, solvate, or prodrug thereof; and
a pharmaceutically appropriate carrier.
52. A method of treating cancer comprising:
administering to a patient in need thereof a therapeutically effective amount of a compound having a formula selected from the group consisting of:
7
or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and a pharmaceutically appropriate carrier.