# US 11,306,074 B2 · Chemical and method for analyzing DNA sequence by visualizing single-molecule DNA and use thereof

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- Number: US 11,306,074 B2
- Document ID: US11306074B2
- Status: Granted
- Kind: B2
- Application number: 16/446,887
- Filed: 2019-06-20
- Published: 2020-01-30
- Granted: 2022-04-19
- Earliest claimed: 2018-07-24
- Assignees: Sogang University Research Foundation
- Applicants: Sogang University Research Foundation
- Inventors: Kyu Bong Jo; Seong Hyun Lee; Hiroshi Sugiyama
- Web page: https://patentgrep.com/patent/US11306074B2

## CPC

- C07D 405/14: containing three or more hetero rings
- C12Q 1/6806: Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence)
- C12Q 1/6816: characterised by the detection means (C12Q1/6804 takes precedence)
- C12Q 2563/107: fluorescence
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present disclosure relates to a composition for analysis of DNA sequences and a method for analysis of DNA sequences by using the same and, more particularly, to a composition comprising the compound represented by Chemical Formula 1 and a method for analysis of DNA sequences, the method comprising a step of treating a sample with the same. The compound represented by Chemical Formula 1 in which TAMRA is linked to polypyrrole specifically binds an A/T base pair (W) to fluoresce alone without causing DNA photocleavage. Therefore, the compound is useful for DNA analysis particularly at the single DNA molecule level.

## Claims

### Claim 1

A composition for analysis of DNA sequences, the composition comprising a compound represented by the following Chemical Formula 1:
wherein,
n is an integer of 2 to 10, m, o, and p are each independently an integer of 1 to 10, and
X is one selected from the group consisting of a fluorescent protein, a photoprotein, a color reaction-catalyst, biotin, a fluorescent substance, a luminescent substance, and a chemiluminescent substance.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the fluorescent substance is one selected from the group consisting of TAMRA (carboxytetramethylrhodamine), fluorescein, Cy5 (Cyanine 5), Cy3 (Cyanine 3), HEX (5′-hexachloro-fluorescein), TET (5′-tetrachloro-fluorescein), Dabsyl (4-(dimethylaminoazo)benzene-4-carboxylic acid), and FAM (fluorescein amidite).

### Claim 3 (depends on 1)

The composition of claim 1, wherein n is an integer of 2 to 5; m, o, and p are each independently an integer of 1 to 5; and
X is

### Claim 4 (depends on 1)

The composition of claim 1, wherein the compound represented by Chemical Formula 1 is a compound represented by the following Chemical Formula 2:

### Claim 5 (depends on 1)

The composition of claim 1, binding to an adenine/thymine (A/T) base pair (W).

### Claim 6 (depends on 1)

The composition of claim 1, wherein the DNA is one selected from the group consisting of a single DNA molecule, a chromosome, and a chromatin fiber.

### Claim 7 (depends on 1)

A method for analysis of DNA sequences comprising:
a step of treating a sample with the composition of claim 1.

### Claim 8 (depends on 7)

The method of claim 7, further comprising:
a step of comparing an entire genomic adenine/thymine (A/T) frequency in a subject to be analyzed and an NT frequency of the sample treated with the composition.

### Claim 9 (depends on 7)

The method of claim 7, wherein the composition binds to an A/T base pair (W).

### Claim 10 (depends on 7)

The method of claim 7, wherein the sample is at least one selected from the group consisting of a single DNA molecule, an oligo-DNA, a chromosome, a polytene chromosome, and a chromatin fiber.