Mapping of genomic interactions

Patent metadata

Field	Value
Publication number	US-9273309-B2
Application number	US-31042707-A
Country	US
Kind code	B2
Filing date	Aug 24, 2007
Priority date	Aug 24, 2006
Publication date	Mar 1, 2016
Grant date	Mar 1, 2016

Abstract

The present invention relates to genomic analysis. In particular, the present invention provides methods and compositions for mapping genomic interactions.

First claim

The invention claimed is:

1. A method, comprising:
a) providing a chromosome conformation capture (3C) library constructed from cross-linked interacting chromatin segments derived from at least one cell, wherein said 3C library comprises a plurality of ligation products representative of interacting genomic loci, wherein each of said plurality of ligation products is formed by two adjacent restriction fragments;
b) annealing unique primer pairs to said plurality of ligation products; and
c) ligating said annealed unique primer pairs together, thereby providing a carbon copy chromosome capture (5C) library.

2. The method of claim 1 , wherein said plurality of unique primer pairs comprises at least 10 unique primer pairs.

3. The method of claim 1 , wherein said plurality of unique primer pairs comprises at least 100 unique primer pairs.

4. The method of claim 1 , wherein said plurality of unique primer pairs comprises at least 500 unique primer pairs.

5. The method of claim 1 , wherein said plurality of unique primers pairs comprises at least 1000 unique primer pairs.

6. The method of claim 1 , wherein said plurality of unique primers pairs comprises at least 10,000 unique primer pairs.

7. The method of claim 1 , wherein said plurality of unique primers pairs comprises at least 100,000 unique primer pairs.

8. The method of claim 1 , wherein said ligation products comprise nucleic acids approximately 100 bps in length.

9. The method of claim 1 , wherein said 5C library is representative of long-range genomic interactions.

10. The method of claim 9 , wherein said long-range genomic interactions comprise interaction of activators or repressors of gene expression with a gene.

11. The method of claim 9 , wherein said long-range genomic interactions comprise interaction of chromatin on different chromosomes.

12. The method of claim 1 , wherein said at least one cell is an animal cell.

13. The method of claim 1 , wherein said at least one cell is selected from the group consisting of a bacterial cell, and a plant cell.

14. The method of claim 9 , further comprising the step of calculating interaction frequencies for said long range genomic interactions.

15. The method of claim 1 , wherein said 3C library has one or more variant genes selected from at least one of the group consisting of polymorphisms, genomic deletions, genomic fusions, genomic translocations, and genomic inversions.

16. The method of claim 1 , further comprising step (d) amplifying said 5C library with a single pair of universal primers.

17. The method of claim 16 , further comprising step (e) analyzing said amplified 5C library with a high-throughput application.

18. A method, comprising:
a) providing a chromosome conformation capture (3C) library constructed from cross-linked interacting chromatin segments derived from at least one cell, wherein said 3C library comprises a plurality of ligated nucleic acid restriction fragments with a junction representative of interacting genomic loci; and
b) performing multiplex ligation mediated amplification with primer pairs targeting said junction.

19. The method of claim 18 , wherein said primer pairs are selected from the group consisting of at least 10 unique primer pairs, at least 100 unique primer pairs, at least 500 unique primer pairs, at least 1000 unique primer pairs, at least 10,000 unique primer pairs, and at least 100,000 unique primer pairs.

20. The method of claim 18 , wherein said ligated restriction fragments comprise nucleic acids approximately 100 bps in length.

21. The method of claim 18 , wherein said multiplex ligation mediated amplification creates a 5C library.

22. The method of claim 21 , wherein said 5C library is amplified with a single pair of universal primers.

23. The method of claim 22 , wherein said amplified 5C library is analyzed with a high-throughput application.

24. The method of claim 23 , wherein said high throughput application comprises sequencing.

25. The method of claim 23 , wherein said high throughput application is performed using a microarray.

26. The method of claim 21 , wherein said 5C library is representative of long-range genomic interactions.

27. The method of claim 26 , wherein said long-range genomic interactions comprise interaction of activators or repressors of gene expression with a gene.

28. The method of claim 26 , wherein said long-range genomic interactions comprise interaction of chromatin on different chromosomes.

29. The method of claim 18 , wherein said at least one cell is an animal cell.

30. The method of claim 18 , wherein said at least one cell is selected from the group consisting of a bacterial cell, and a plant cell.

31. The method of claim 26 , further comprising the step of calculating interaction frequencies for said long range genomic interactions.

32. The method of claim 18 , wherein said 3C library has one or more variant genes selected from at least one of the group consisting of polymorphisms, genomic deletions, genomic fusions, genomic translocations, and genomic inversions.

33. The method of claim 17 , wherein said high throughput application comprises sequencing.

34. The method of claim 17 , wherein said high throughput application is performed using a microarray.

Assignees

• Dekker Job
• Dostie Josee
• Univ Massachusetts

Inventors

• Dekker Job
• Dostie Josee

Classifications

• C12N15/1093

  General methods of preparing gene libraries, not provided for in other subgroups · CPC title

• C12N15/1072 Primary

  Differential gene expression library synthesis, e.g. subtracted libraries, differential screening · CPC title