Novel methods for high-throughput genome-wide location analysis
First Claim
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1. A method for identifying a region of a genome of a cell to which a protein of interest is bound, the method comprising the steps of:
- (a) fragmenting the genomic DNA of the cell by;
(i) a mechanical or chemical process; and
(ii) by an enzymatic means, thereby producing a mixture comprising DNA fragments to which the protein of interest is bound;
(b) isolating a DNA fragment to which the protein of interest is bound from the mixture produced in step (a); and
(c) identifying a region of the genome of the cell which is complementary to the DNA fragment isolated in step (b), thereby identifying a region of a genome of a cell to which the protein of interest is bound.
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Abstract
The invention relates to improved methods of identifying the genomic regions to which a protein of interest binds, and in particular, to methods that are highly-sensitive and/or high throughput. The invention also provides methods of identifying agents which modulate the binding of a protein to the genome of a cell and methods of identifying variant proteins, such as transcription factors, with altered genome-binding properties. The invention also provides kits related to the methods described herein.
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Citations
85 Claims
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1. A method for identifying a region of a genome of a cell to which a protein of interest is bound, the method comprising the steps of:
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(a) fragmenting the genomic DNA of the cell by;
(i) a mechanical or chemical process; and
(ii) by an enzymatic means, thereby producing a mixture comprising DNA fragments to which the protein of interest is bound;
(b) isolating a DNA fragment to which the protein of interest is bound from the mixture produced in step (a); and
(c) identifying a region of the genome of the cell which is complementary to the DNA fragment isolated in step (b), thereby identifying a region of a genome of a cell to which the protein of interest is bound. - View Dependent Claims (2, 3, 19, 45, 51, 52, 54, 55, 56, 57, 58, 63, 65, 74, 76, 77)
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4-18. -18. (canceled)
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20-21. -21. (canceled)
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23-44. -44. (canceled)
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46-50. -50. (canceled)
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59-62. -62. (canceled)
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64. (canceled)
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66-73. -73. (canceled)
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75. (canceled)
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78-83. -83. (canceled)
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84. A kit comprising (i) one or more DNA endonucleases;
- and (ii) a dilution buffer lacking free amino groups.
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85-93. -93. (canceled)
Specification