IP Library Granted Patent US 9,434,985
Granted Patent B2
US 9,434,985 · App. 13/121,158 · Granted Sep 6, 2016

Methods of identifying interactions between genomic loci

Inventors: Job Dekker (Princeton, MA); Erez Lieberman (Cambridge, MA); Nynke Van Berkum (The Hague, NL); Andreas Gnirke (Wellesley, MA); Eric Lander (Cambridge, MA); Chad Nusbaum (Nowtoa, MA); Louise Williams (Reading, MA); Alexandre Melnikov (Bellingham, MA); Georgia Giannoukos (Cambridge, MA)
Assignees: University of Massachusetts; Massachusetts Institute Of Technology; President And Fellows Of Harvard College; Whitehead Institute For Biomedical Research
C12Q1/6837C12Q1/68
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Quick Facts
Patent No.
US 9,434,985
App. No.
13/121,158
Granted
Sep 6, 2016
Kind
B2
Abstract

The disclosed Hi-C protocol can identify genomic loci that are spatially co-located in vivo. These spatial co-locations may include, but are not limited to, intrachromosomal interactions and/or interchromosomal interactions. Hi-C techniques may be applied to many different scales of interest. For example, on a large scale, Hi-C techniques can be used to identify long-range interactions between distant genomic loci.

Claims (50)

1. A method for identifying interaction frequencies, comprising:

a) providing;

i) a nuclear matrix comprising a first genomic region and a second genomic region; and

ii) a junction marker labeled with an affinity marker for selective purification of a ligation product;

b) fragmenting said first genomic region and said second genomic region into a plurality of first genomic region fragments and a plurality of second genomic region fragments;

c) ligating said junction marker between at least one of said first genomic region fragments and at least one of said second genomic region fragments to create said ligation product;

d) purifying said ligation product with said affinity marker;

and

e) analyzing said ligation product under conditions such that a genomic interaction frequency for said ligation product is identified.

2. The method of claim 1 , wherein said affinity marker comprises biotin.

3. The method of claim 1 , wherein said first and second genomic regions are located on the same chromosome.

4. The method of claim 1 , wherein said first and second genomic regions are located on different chromosomes.

5. The method of claim 1 , wherein said interaction frequency identifies a long range interaction.

6. The method of claim 1 , wherein said interaction frequency identifies a short range interaction.

7. The method of claim 1 , wherein said interaction frequency identifies a close neighbor interaction.

8. The method of claim 1 , wherein said nuclear matrix is derived from a human cell nucleus.

9. The method of claim 1 , wherein said nuclear matrix is derived from a yeast cell nucleus.

10. The method of claim 1 , further comprising contacting said junction marker with a ligand.

11. The method of claim 10 , wherein said ligand comprises streptavidin.

12. The method of claim 1 , further comprising digesting said nuclear matrix fragments with a restriction endonuclease wherein a first region fragment comprising a first sticky end and a second region fragment comprising a second sticky end are created.

13. The method of claim 12 , further comprising filling in said first sticky end and said second sticky end to create a first blunt end and a second blunt end; and ligating said first and second blunt ends, wherein said first region fragment and said second region fragment are joined.

14. The method of claim 1 , further comprising digesting said nuclear matrix fragments with an exonuclease.

15. A method for identifying interaction frequencies, comprising:

a) crosslinking a nuclear matrix comprising a first genomic region and a second genomic region;

b) fragmenting said first genomic region and said second genomic region into a plurality of first genomic region fragments and a plurality of second genomic region fragments;

c) ligating a junction marker labeled with an affinity marker between at least one of said first genomic region fragments and at least one of said second genomic region fragments to create a ligation product;

d) purifying said ligation product with said affinity marker;

and

e) analyzing said ligation product under conditions such that a genomic interaction frequency for said ligation product is identified.

16. A method for identifying interaction frequencies, comprising:

a) crosslinking a nuclear matrix comprising a first genomic region and a second genomic region;

b) fragmenting said first genomic region and said second genomic region into a plurality of first genomic region fragments and a plurality of second genomic region fragments;

c) ligating a junction marker labeled with an affinity marker between at least one of said first genomic region fragments and at least one of said second genomic region fragments to create a ligation product;

d) purifying said ligation product with said affinity marker;

and

e) detecting a close proximity between said first genomic region and said second genomic region with said ligation product under conditions such that a genomic interaction frequency for said ligation product is identified.

17. A method for identifying interaction frequencies, comprising:

a) crosslinking a nuclear matrix comprising a first genomic region and a second genomic region;

b) fragmenting said first genomic region and said second genomic region into a plurality of first genomic region fragments and a plurality of second genomic region fragments;

c) ligating a junction marker labeled with an affinity marker between at least one of said first genomic region fragments and at least one of said second genomic region fragments to create a ligation product;

d) purifying said ligation product with said affinity marker;

and

e) identifying a contact probability as a function of genomic distance between said first genomic region and said second genomic region with said ligation product under conditions such that a genomic interaction frequency for said ligation product is identified.

18. A method for identifying interaction frequencies, comprising:

a) crosslinking a nuclear matrix comprising a first genomic region and a second genomic region;

b) fragmenting said first genomic region and said second genomic region into a plurality of first genomic region fragments and a plurality of second genomic region fragments;

c) ligating a junction marker labeled with an affinity marker between at least one of said first genomic region fragments and at least one of said second genomic region fragments to create a ligation product;

d) purifying said ligation product with said affinity marker;

and

e) identifying whether said first genomic region and said second genomic region preferentially associate with each other with said ligation product under conditions such that a genomic interaction frequency for said ligation product is identified.

Assignments (6)
CONFIRMATORY LICENSE Recorded May 28, 2015
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 035786/0319 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 4, 2012
From: LIEBERMAN, EREZ
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY; PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Reel/Frame 027475/0490 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 3, 2011
From: DEKKER, JOB; VAN BERKUM, NYNKE
To: UNIVERSITY OF MASSACHUSETTS
Reel/Frame 027005/0866 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 3, 2011
From: GNIRKE, ANDREAS; NUSBAUM, CHAD; WILLIAMS, LOUISE; MELNIKOV, ALEXANDRE; GIANNOUKOS, GEORGIA
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 027006/0294 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 3, 2011
From: LIEBERMAN-AIDEN, EREZ
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY; PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Reel/Frame 027006/0939 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 3, 2011
From: LANDER, ERIC
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY; PRESIDENT AND FELLOWS OF HARVARD COLLEGE; WHITEHEAD INSTITUTE FOR BIOMEDICAL RESEARCH
Reel/Frame 027007/0636 →
Continuity (2)
Provisional Application 61100151 · Sep 25, 2008
Related Publication 20130096009A1 · Apr 18, 2013