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US 2019/0292580 A1Published application
Dna logic-gated proximity assembly circuit for biochemical sensing
AssigneeRutgers, The State University of New Jersey
InventorsJinglin Fu, Sung Won Oh, Adriana Rosa Pereira
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Abstract
Provided are nucleic acid-based sensors, kits that include such sensors, and methods for making and using such sensors. The sensors permit detection of a broad array of target agents, such as nucleic acids (e.g., DNA and RNA), proteins, cells, and small molecules (e.g., toxins and metals). In some examples, the sensors determine the activity of a protein, such as an enzyme's activity. The assays provide easy-to-read signals for point-of-care diagnosis. The sensors control the conformational switch for recognizing targets and to trigger the subsequent assembly of enzyme/cofactor structures for producing and amplifying signals.
Claims22
A sensor, comprising:
- (a) a trigger module, comprising
- (i) a first nucleic acid molecule, comprising a first portion specific for a target, a second portion, and a third portion,
- (ii) a second nucleic acid molecule comprising a terminal cofactor, a first stem portion, a loop portion, a second stem portion, a third stem portion, and a fourth portion,
- wherein the first stem portion includes the terminal cofactor and is complementary to the second stem portion, thereby resulting in hybridization between the first stem portion and the second stem portion, thereby producing the loop portion,
- wherein the third stem portion is complementary to the second portion of the first nucleic acid molecule, thereby resulting in hybridization between the third stem portion and the second portion of the first nucleic acid molecule,
- (iii) a third nucleic acid molecule, comprising a first portion and second portion,
- wherein the first portion of the third nucleic acid molecule is complementary to the third portion of the first nucleic acid molecule thereby resulting in hybridization between the first portion of the third nucleic acid molecule and the third portion of the first nucleic acid molecule,
- wherein the second portion of the third nucleic acid molecule is complementary to the fourth portion of the second nucleic acid molecule thereby resulting in hybridization between the second portion of the third nucleic acid molecule and the fourth portion of the second nucleic acid molecule,
- wherein hybridization between nucleotides of the first and second nucleic acid molecules, first and third nucleic acid molecules, and second and third nucleic acid molecules, forms the trigger module,
- (b) sensory module, comprising
- (i) a fourth nucleic acid molecule comprising a nucleic acid strand;
- and
- (ii) an enzyme,
- wherein the nucleic acid strand of the fourth nucleic acid molecule is complementary to the second and third stem portion of the second nucleic acid molecule, and wherein the enzyme is activated by the cofactor.
11 more elements
A sensor, comprising:
- (a) a trigger module, comprising
- (i) a first nucleic acid molecule, comprising a first portion specific for a target and a second portion,
- (ii) a second nucleic acid molecule comprising a terminal cofactor, a first stem portion, a loop portion, a second stem portion, and a third stem portion, wherein the first stem portion includes the terminal cofactor and is complementary to the second stem portion, thereby resulting in hybridization between the first stem portion and the second stem portion, thereby producing the loop portion,
- wherein the third stem portion is complementary to the second portion of the first nucleic acid molecule, thereby resulting in hybridization between the third stem portion and the second portion of the first nucleic acid molecule,
- wherein hybridization between nucleotides of the first and second nucleic acid molecules, forms the trigger module,
- (b) sensory module, comprising
- (i) a third nucleic acid molecule comprising a nucleic acid strand;
- and
- (ii) an enzyme,
- wherein the nucleic acid strand of the third nucleic acid molecule is complementary to the second and third stem portion of the second nucleic acid molecule, and wherein the enzyme is activated by the cofactor.
7 more elements
A sensor, comprising:
- (a) a sensory module, comprising
- (i) a first nucleic acid molecule comprising a nucleic acid strand;
- and
- (ii) a terminal enzyme, and
- (b) a trigger module, comprising
- (i) a second nucleic acid molecule comprising loop portion and an aptamer portion, and
- (ii) a terminal inhibitor of the enzyme,
- wherein at least a portion of the aptamer portion is complementary to the first nucleic acid molecule, thereby resulting in hybridization between the first nucleic acid molecule of the sensor module and the aptamer portion of the trigger module, thereby bringing the inhibitor of the enzyme and the enzyme into proximity such that the inhibitor of the enzyme inhibits the enzyme.
5 more elements
Dependencies are inferred from the claim text.