Molecular Switches And Methods For Their Use
AssigneeJohnson & Johnson Research Pty. Limited
InventorsAlison V. Todd, Elisa Mokany, Donald J. Birkett, Tram Bich Doan, Christopher R. Reid
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Abstract
The present invention relates to compositions and methods that allow the manipulation of the catalytic activity of multi-component nucleic acid complexes. Further, the invention provides methods which use these compositions and methods to create molecular sensors, molecular switches, and/or modulators or propagators of autocatalytic self-replicating cascades and other iterative processes. More particularly, the invention relates to compositions allowing self-assembly of active and inactive multicomponent nucleic acid complexes, methods of making such compositions, and methods for use.
Claims20
1-110. (canceled)
A molecular switch comprising at least one oligonucleotide partzyme, wherein said partzyme comprises at least one of a catalytic core portion, a sensor arm portion, and a substrate arm portion, and at least one further oligonucleotide component selected from the group comprising an assembly facilitator oligonucleotide, a substrate oligonucleotide, an activity inhibitor and an assembly inhibitor or any combination thereof wherein said molecular switch is capable of forming a multicomponent nucleic acid (MNA) complex.
Use of an MNA complex as a molecular switch, wherein said complex transitions from an active to an inactive MNA complex or from an inactive to an active MNA in response to an input event.
A method of detecting a target using a cascade, wherein said cascade comprises an initiating MNAzyme formed in the presence of said target;
- a first MNAzyme formed in the presence of a product of said initiating MNAzyme wherein said method comprises the steps of;
- (i) modifying a first substrate with said initiating MNAzyme to generate a first assembly facilitator;
- (ii) assembling said first MNAzyme with said first assembly facilitator;
- (iii) modifying an additional substrate with said first MNAzyme and
- wherein said modification of at least one of said first or said additional substrates produces a detectable effect indicative of the presence of said target.
A method of detecting a target using a cascade, wherein said casade comprises an initiating MNAzyme formed in the presence of said target;
- a first MNAzyme formed in the presence of a product of said initiating MNAzyme;
- an additional MNAzyme formed in the presence of a product of said first MNAzyme wherein said method comprises the steps of;
- (i) modifying a first substrate with said initiating MNAzyme to generate a first assembly facilitator;
- (ii) assembling said first MNAzyme with said first assembly facilitator;
- (iii) modifying an additional substrate with said first MNAzyme to generate an additional assembly facilitator;
- (iv) assembling said additional MNAzyme with said additional assembly facilitator;
- (v) modifying said first substrate with said additional MNAzyme to generate said first assembly facilitator;
- (vi) assembling said first MNAzyme with said first assembly facilitator released from (v) thereby forming an amplification cascade;
- and
- wherein said modification of at least one of said first or said additional substrates produces a detectable effect indicative of the presence of said target.
An MNAi comprising two or more oligonucleotide components and at least one activity inhibitor molecule.
An MNAi comprising at least two or more oligonucleotide components wherein at least a first oligonucleotide component and a second oligonucleotide component are capable of self-assembly in the presence of at least one MNA complex activity inhibitor wherein each of said at least first and said second oligonucleotide components comprise a substrate arm portion, a catalytic core portion, and a sensor arm portion;
- wherein upon self-assembly, the sensor arm portion of said first and second oligonucleotide components act as sensor arms, the substrate arm portion of the first and second oligonucleotide components act as substrate arms, and the catalytic core portion of the first and second oligonucleotide components form the non-functional catalytic core;
- and
- wherein upon self-assembly at least one sensor arm interacts with said activity inhibitor and said first and second oligonucleotide components are maintained in proximity for association of their respective catalytic core portions to form a non-functional catalytic core.
A method for the detection of an assembly facilitator using a signal cascade comprising a first MNAzyme, an MNA complex initially present in a substantially catalytically inactive form (MNAi), an activity inhibitor capable of being modified by said first MNAzyme to provide a detectable effect, wherein said activity inhibitor is both an activity inhibitor and a potential substrate;
- and
- wherein association of said assembly facilitator with partzymes for said first MNAzyme under conditions permitting catalytic activity of said first MNAzyme facilitates the catalytic activity of said first MNAzyme thereby providing modification of said activity inhibitor to release an activator assembly facilitator domain and an activity inhibitor domain of said activity inhibitor and wherein said release provides said detectable effect;
- and
- wherein said released activator assembly facilitator domain facilitates assembly of a second MNAzyme from components of said MNA complex;
- and
- wherein catalytic activity of said second MNAzyme modifies said activity inhibitor to release further activity inhibitor domains and further activator assembly facilitator domains, and wherein said release provides further detectable effect, and;
- wherein said further activator assembly facilitator domains facilitate assembly of additional second MNAzymes thereby providing further said catalytically active second MNAzymes thereby providing further detectable effect indicative of the presence of said assembly facilitator, and;
- wherein the assembly of catalytically active MNAzymes is regulated by an input event selected from the group comprising change in temperature, salt concentration, ionic strength, pH, divalent cation presence or absence, type or concentration, electric charge, magnetic charge, physical manipulation and change in concentration of an MNA or modulator component or component of the microenvironment, or any combination thereof.
Dependencies are inferred from the claim text.