Claims for Patent: 7,700,284

Summary for Patent: 7,700,284

Title:	Reporter transcription unit populations and kits comprising same
Abstract:	Compositions, methods and kits are provided that are useful, for example, for determining activities of multiple cis-regulatory sequences, such as promoters and enhancers, and/or multiple trans-acting factors, such as transcription factors, in a cell. In particular, in certain embodiments, compositions are provided comprising a population of polynucleotide reporter transcription units (RTUs) in which each RTU comprises a reporter sequence, a processing tag located in the reporter sequence; and a cis-regulatory element operably linked to the reporter sequence, wherein the reporter sequences between any two RTUs in the population, outside of the processing tags, are substantially identical and wherein the positions of the processing tags within the reporter sequences distinguish between any two RTUs differing, for example, in their cis-regulatory elements. The compositions, methods and kits can further be used, for example, to identify a cell type or disease state, for example, in a biological organism.
Inventor(s):	Romanov; Sergei R. (Chapel Hill, NC), Medvedev; Alex (Durham, NC), Makarov; Sergei S. (Chapel Hill, NC)
Assignee:	Attagene, Inc. (Morrisville, NC)
Application Number:	11/271,190
Patent Claims:	1. A reporter transcription unit (RTU) population comprising polynucleotide RTUs, each RTU comprising a reporter sequence; a processing tag located in the reporter sequence; and a cis-regulatory element operably linked to the reporter sequence, wherein transcription of the reporter sequence is modulated by the cis-regulatory element; and wherein the reporter sequences of any two RTUs in the population, outside of the processing tags, are identical or substantially identical, wherein the processing tags are identical between any two RTUs in the population, and wherein the reporter sequences of any two RTUs operably linked to non-identical cis-regulatory elements are distinguishable by the locations of the processing tags in the reporter sequences. 2. The RTU population of claim 1, wherein a trans-acting factor binds to at least one cis-regulatory element. 3. The RTU population of claim 1, wherein the reporter sequences between any two RTUs in the population, outside of the processing tags, are substantially identical. 4. The RTU population of claim 3, wherein the substantially identical reporter sequences differ by less than 10 nucleotides. 5. The RTU population of claim 3, wherein the substantially identical reporter sequences differ by 1 nucleotide. 6. The RTU population of claim 1, wherein the reporter sequences, outside of the processing tags, are identical between any two RTUs in the population. 7. The RTU population of claim 1, wherein the reporter sequences, including the processing tags, between any two RTUs in the population comprise an identical number of nucleotides. 8. The RTU population of claim 7, wherein the positions of the processing tags in the reporter sequences differ by at least 1 to 15 nucleotides between any two RTUs in the population. 9. The RTU population of claim 8, wherein the reporter sequences, outside of the processing tags, are identical between any two RTUs in the population. 10. The RTU population of claim 1, wherein the processing tag for each RTU in the population is selected from the group consisting of: a) a thymine, adenine, cytosine, or guanine nucleotide residue; b) an endonuclease recognition site; c) a primer sequence; d) a polyadenylation termination signal; and e) a mutation in the reporter sequence comprising a deletion, insertion, or substitution. 11. The RTU population of claim 10, wherein the processing tag is selected from the group consisting of: a thymine, adenine, cytosine, or guanine nucleotide residue; and an endonuclease recognition site. 12. The RTU population of claim 1, wherein at least one of the RTUs in the population comprises a cis-regulatory element selected from the group consisting of a promoter, an enhancer, an RNA stability signal, and a polyadenylation signal or a combination thereof. 13. The RTU population of claim 1, wherein, for each RTU in the population, the cis-regulatory element comprises a promoter or an enhancer, and has at least one binding site for a DNA-binding protein. 14. The RTU population of claim 13, wherein the binding site is a transcription factor binding site. 15. The RTU population of claim 13, wherein the cis-regulatory element modulates the stability or maturation of the reporter species. 16. The RTU population of claim 1, wherein each RTU in the population further comprises a 5' and a 3' primer sequence that flank the reporter sequence. 17. The RTU population of claim 1, wherein at least one of the RTUs in the population comprises an intron. 18. A reporter transcription unit (RTU) population comprising polynucleotide RTUs, each RTU comprising a reporter sequence; a processing tag located in the reporter sequence; and a cis-regulatory element operably linked to the reporter sequence, wherein transcription of the reporter sequence is modulated by binding of a trans-acting factor to the cis-regulatory element; and wherein the reporter sequences of any two RTUs in the population, outside of the processing tags, are identical or substantially identical, wherein the processing tags are identical between any two RTUs in the population, and wherein the reporter sequences of any two RTUs operably linked to cis-regulatory elements modulated by non-identical trans-acting factors are distinguishable by the locations of the processing tags in the reporter sequences. 19. A population of vectors comprising the RTU population of claim 1. 20. The population of vectors of claim 19, wherein each vector is selected from the group consisting of a plasmid, a phagemid, a retroviral vector, a lentiviral vector, an adenoviral vector, and an adeno-associated vector. 21. A cell comprising the RTU population of claim 1. 22. The cell of claim 21, wherein the RTU population is stably incorporated into the genome of said cell. 23. The cell of claim 21, wherein the cell is selected from the group consisting of a plant, a bacterium, a fungus, and a vertebrate. 24. The cell of claim 21, wherein the cell is an embryonic stem cell. 25. A tissue comprising the cell of claim 21. 26. The tissue of claim 25, wherein said tissue comprises a biopsy sample, an autopsy sample, or a slice tissue culture. 27. A population of cells comprising the RTU population of claim 1, wherein each cell of the cell population comprises at least one RTU member of the RTU population. 28. A population of cells comprising the RTU population of claim 1, wherein each cell of the cell population comprises each member of the RTU population. 29. A non-human organism comprising the cell of claim 21. 30. A kit comprising the RTU population of claim 1. 31. A kit comprising a cell, wherein the cell comprises the RTU population of claim 1. 32. A kit comprising a vector population, wherein the vector population comprises the RTU population of claim 1. 33. A tissue comprising the population of cells of claim 27. 34. A tissue comprising the population of cells of claim 28. 35. A non-human organism comprising the population of cells of claim 27. 36. A non-human organism comprising the population of cells of claim 28.

Details for Patent 7,700,284

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		2024-11-10
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2024-11-10
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2024-11-10