# US 7,612,185 B2 · Nucleic acid biosensors

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- Number: US 7,612,185 B2
- Document ID: US7612185B2
- Status: Granted
- Kind: B2
- Application number: 10/384,497
- Filed: 2003-03-07
- Published: 2004-09-09
- Granted: 2009-11-03
- Earliest claimed: 2003-03-07
- Assignees: The Board of Trustees of the University of Illinois
- Inventors: Yi Lu; Juewen Liu
- Web page: https://patentgrep.com/patent/US7612185B2

## CPC

- C12Q 1/25: involving enzymes not classifiable in groups C12Q1/26 {- C12Q1/66}
- C12Q 1/68: involving nucleic acids
- 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

Sensors comprising aptazymes capable of detecting the presence and concentration of effectors, as well as methods of using such sensors, are disclosed.

## Claims

### Claim 1

A sensor system for detecting an effector, comprising:
(i) an aggregate, wherein the aggregate comprises a plurality of complexes wherein each complex comprises:
(I) an aptazyme, comprising an aptamer comprising a binding site for the effector,
(II) a substrate for the aptazyme, hybridized to the aptazyme,
(III) particles,
(IV) polynucleotides attached to the particles at the 5′ terminus or 3′ terminus and hybridized to the substrate,
wherein the aptazyme cleaves the substrate when the aptamer binds the effector and cleavage of the substrate results in deaggregation of the aggregate.

### Claim 2

A sensor system for detecting an effector, comprising:
(i) an aggregate, wherein the aggregate comprises a plurality of complexes wherein each complex comprises:
(I) an aptazyme, comprising an aptamer comprising a binding site for the effector,
(II) a substrate for the aptazyme, hybridized to the aptazyme,
(III) gold particles,
(IV) polynucleotides attached to the particles at the 5′-terminus or 3′-terminus, and hybridized to the substrate, and
(ii) Mg(II),
wherein the aptazyme comprises DNA,
the aptazyme cleaves the substrate when the aptamer binds the effector and cleavage of the substrate results in deaggregation of the aggregate.

### Claim 3

A sensor system for detecting an effector, comprising:
(i) an aggregate, wherein the aggregate comprises a plurality of complexes wherein each complex comprises:
(I) an aptazyme, comprising an aptamer comprising a binding site for the effector,
(II) a substrate for the aptazyme, hybridized to the aptazyme,
(III) first particles,
(IV) first polynucleotides attached to the first particles at the 5′-terminus or 3′-terminus and hybridized to the substrate;
(V) second particles,
(VI) second polynucleotides attached to the second particles at the 5′-terminus or 3′-terminus and hybridized to the substrate,
wherein the aptazyme cleaves the substrate when the aptamer binds the effector and cleavage of the substrate results in deaggregation of the aggregate.

### Claim 4 (depends on 1)

The sensor system of claim 1, wherein the aptazyme comprises DNA.

### Claim 5 (depends on 1)

The sensor system of claim 1, wherein the effector is adenosine.

### Claim 6 (depends on 1)

The sensor system of claim 1, wherein the particles comprise gold particles.

### Claim 7 (depends on 1)

The sensor system of claim 1, wherein the particles comprise a material selected from the group consisting of metal colloids, semiconductor colloids and polystyrene latex particles.

### Claim 8 (depends on 2)

The sensor system of claim 2, further comprising Pb(II).

### Claim 9 (depends on 3)

The sensor system of claim 3, wherein the aptazyme comprises DNA.

### Claim 10 (depends on 3)

The sensor system of claim 3, wherein the effector is adenosine.

### Claim 11 (depends on 3)

The sensor system of claim 3, wherein the first and second particles comprise gold particles.

### Claim 12 (depends on 3)

The sensor system of claim 3, wherein the first and second particles comprise a material selected from the group consisting of metal colloids, semiconductor colloids and polystyrene latex particles.

### Claim 13 (depends on 3)

The sensor system of claim 3, wherein the first and second polynucleotides have identical sequences.

### Claim 14 (depends on 3)

The sensor system of claim 3, wherein the first and second polynucleotides have different sequences.

### Claim 15 (depends on 1)

The sensor system of claim 1, wherein the effector activates the aptazyme.

### Claim 16 (depends on 1)

The sensor system of claim 1, wherein the effector inhibits the aptazyme.

### Claim 17 (depends on 2)

The sensor system of claim 2, wherein the effector activates the aptazyme.

### Claim 18 (depends on 2)

The sensor system of claim 2, wherein the effector inhibits the aptazyme.

### Claim 19 (depends on 3)

The sensor system of claim 3, wherein the effector activates the aptazyme.

### Claim 20 (depends on 3)

The sensor system of claim 3, wherein the effector inhibits the aptazyme.