Method of determining chromatin structure
AssigneeImperial College Innovations Limited
InventorRobert Otto Johannes Weinzierl
Life of this patent
Estimating the term…
Abstract
A method of determining chromatin structure is described. The method comprises the steps of (i) fragmenting a nucleotide sequence at multiple HSs; and (ii) analysing fragments formed in step (i) from a plurality of sequences. In a preferred aspect, the present invention provides a method of determining chromatin structure in a nucleic acid sample comprising the steps of (i) treating the sample to fragment the nucleic acid therein at multiple HSs; and (ii) analysing fragments formed in step (i) from a plurality of genes.
Claims19
A method of determining chromatin structure of a nucleic acid comprising the steps of (i) fragmenting a nucleic acid sample comprising a nucleosome at multiple hypersensitive sites (HSs) using an enzyme, chemical agent or physical agent;
- (ii) creating a hypersensitive site library comprising the fragments formed from at least 10 genes in step (i);
- and (iii) analyzing fragments from a plurality of genes contained in the library to identify the location of a plurality of HS sites.
2-3. (canceled)
A method of determining chromatin structure in a nucleic acid sample comprising the steps of (i) treating a nucleic acid sample comprising a nucleosome with a first restriction enzyme capable of fragmenting the nucleic acid at HSs;
- (ii) incorporating detectable nucleotides into the fragments formed from at least 10 genes in step (i);
- (iii) isolating the nucleic acid from the sample of step (ii);
- (iv) treating the nucleic acid from step (iii) with a second restriction enzyme;
- (v) creating a hypersensitive site library comprising the fragments from step (iv);
- and (vi) analysing the fragments from step (v) from a plurality of genes.
A method of determining chromatin structure in a nucleic acid sample comprising the steps of (i) treating the sample with a first restriction enzyme capable of fragmenting the nucleic acid at multiple HSs;
- (ii) incorporating a target nucleotide sequence into the fragments formed in step (i);
- (iii) isolating the nucleic acid;
- (iv) selectively amplifying portions of the nucleic acid from step (iii) using a first primer capable of binding to the target nucleotide sequence and a second primer capable of binding to another portion of the nucleic acid;
- and (v) analysing the amplified portions from step (iv).
A method of determining a characteristic of a nucleic acid sample comprising the steps of (i) treating a nucleic acid sample comprising a nucleosome to fragment the nucleic acid therein at hypersensitive sites (HSs) using an enzyme, chemical agent or physical agent;
- (ii) creating a hypersensitive site library comprising the fragments formed from at least 10 genes in step (i);
- (iii) analysing fragments from a plurality of genes contained in the library to determine their presence or absence;
- and (iv) using the pattern of fragments from step (iii) to determine the characteristic.
A method of diagnosing a disease in subject wherein the disease is associated with an altered chromatin structure the method comprising the steps of (i) treating a nucleic acid sample from the subject to fragment the nucleic acid therein at multiple HSs;
- (ii) analysing fragments formed in step (i) from a plurality of genes;
- and (iii) using the fragment pattern to diagnose the disease.
A method of monitoring the progress of a disease in a subject, the method comprising the steps of (i) treating a nucleic acid sample from the subject to fragment the nucleic acid therein at multiple HSs;
- (ii) analysing fragments formed in step (i) from a plurality of genes;
- and (iii) using the pattern of fragments from step (ii) to determine the stage of the disease.
A method of testing the efficacy of a putative therapeutic agent which may be suitable for treating a disease in a subject associated with an altered chromatin structure comprising using said agent in a method according to any one of the preceding claims and then determining if said agent is capable of modulating said HS.
14-23. (canceled)
A method of preparing a HS library comprising the steps of (i) treating a nucleic acid sample to fragment the nucleic acid therein at multiple HSs;
- (ii) ligating the nucleic acid fragments formed in step (i) from a plurality of sequences in to a vector;
- and optionally (iii) transforming the vector into a host cell to provide a library of cells.
A library of HSs wherein each library entry comprises HSs from a plurality of genes.
(canceled)
Dependencies are inferred from the claim text.