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US 8,710,199 B2Granted
Signal activated molecular delivery
AssigneesCalifornia Institute of Technology, City of Hope
InventorsSi-Ping Han, William A. Goddard, III, Lisa Scherer, John J. Rossi
Life of this patent
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Abstract
Provided herein are signal activatable molecular constructs for enzyme-assisted delivery of molecules and related components, such as a sensor domain, compositions, methods and systems.
Claims33
A sensor domain for enzyme-assisted molecular delivery, the sensor domain comprising
- a protection segment comprising an RNA portion, and a targeting domain binding portion presented for covalent attachment to a targeting domain;
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide,
- wherein the protection segment, displacement segment, and activation segment are configured so that upon complementary binding of the signal polynucleotide to the toehold segment, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding;
- wherein the targeting domain binding portion and the RNA portion of the protection segment are configured to allow release of the targeting domain upon cleavage of the RNAase H binding site;
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ of the activation segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the activation segment is adjacent to the 5′ terminus of the displacement segment, the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the toehold segment.
6 more elements
A molecular complex for enzyme-assisted molecular delivery, the molecular complex comprising:
- a targeting domain;
- a sensor domain, the sensor domain comprising
- a protection segment comprising an RNA portion,
- an activation segment comprising a DNA portion complementary to the RNA
- portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide wherein the sensor domain is bound to the targeting domain through covalent attachment of the targeting domain to the protection segment of the sensor domain at a targeting domain binding portion configured to allow release of the targeting domain upon cleavage of the RNAase H binding site;
- wherein the protection segment, displacement segment, and activation segment are configured so that upon binding of the signal polynucleotide, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding;
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ of the activation segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the activation segment is adjacent to the 5′ terminus of the displacement segment, the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the toehold segment.
8 more elements
An activatable molecular complex comprising
- a targeting domain and
- a sensor domain, the sensor domain comprising
- a protection segment comprising an RNA portion;
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide the activatable molecular complex configured to exhibit a first conformation and a second, activated, conformation wherein,
- in the first conformation the protection segment covalently attaches the targeting domain, the displacement segment complementarily binds the RNA portion of the protection segment, and the toehold segment and at least a portion of the activation segment are presented for binding to complementary molecules;
- in the second, activated, conformation the displacement segment and the toehold segment complementary bind the signal polynucleotide, the RNA portion of the protection segment complementary binds the DNA portion of the activation segment to provide an RNAase H binding site presented for binding, and the targeting domain is attached to the protection segment in a configuration cleavable upon cleavage of the RNAase binding site;
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ of the activation segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the activation segment is adjacent to the 5′ terminus of the displacement segment, the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the toehold segment.
8 more elements
An activated molecular complex, the activated molecular complex comprising
- a targeting domain;
- and
- a sensor domain, the sensor domain comprising
- a protection segment comprising an RNA portion;
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide
- wherein the displacement segment and the toehold segment complementary bind the signal polynucleotide, the RNA portion of the protection segment complementary binds the DNA portion of the activation segment to provide an RNAase H binding site presented for binding, and the targeting domain is attached to the protection segment in a configuration cleavable upon cleavage of the RNAase binding site
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ of the activation segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the activation segment is adjacent to the 5′ terminus of the displacement segment, the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the toehold segment.
8 more elements
A sensor domain for enzyme-assisted molecular delivery, the sensor domain comprising
- a protection segment comprising an RNA portion, and a targeting domain binding portion presented for covalent attachment to a targeting domain;
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide, wherein the protection segment, displacement segment, and activation segment are configured so that upon complementary binding of the signal polynucleotide to the toehold segment, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding;
- wherein the targeting domain binding portion and the RNA portion of the protection segment are configured to allow release of the targeting domain upon cleavage of the RNAase H binding site, and
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and wherein the 5′ terminus of the activation segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the activation segment is adjacent to the 5′ terminus of the toehold segment, and the 3′ terminus of the toehold segment is adjacent to the 5′ terminus of the displacement segment.
4 more elements
A sensor domain for enzyme-assisted molecular delivery, the sensor domain comprising
- a protection segment comprising an RNA portion, and a targeting domain binding portion presented for covalent attachment to a targeting domain;
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide, wherein the protection segment, displacement segment, and activation segment are configured so that upon complementary binding of the signal polynucleotide to the toehold segment, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding;
- wherein the targeting domain binding portion and the RNA portion of the protection segment are configured to allow release of the targeting domain upon cleavage of the RNAase H binding site,
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ terminus of the toehold segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the toehold segment is adjacent to the 5′ terminus of the displacement segment, and the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the activation segment.
5 more elements
A sensor domain for enzyme-assisted molecular delivery, the sensor domain comprising
- a protection segment comprising an RNA portion, and a targeting domain binding portion presented for covalent attachment to a targeting domain;
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide,
- wherein the protection segment, displacement segment, and activation segment are configured so that upon complementary binding of the signal polynucleotide to the toehold segment, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding;
- wherein the targeting domain binding portion and the RNA portion of the protection segment are configured to allow release of the targeting domain upon cleavage of the RNAase H binding site,
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ terminus of the activation segment is adjacent to the 3′ terminus of the protection segment, the 3′ terminus of the activation segment is arranged as a single strand overhang presented for binding, and the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the toehold segment.
6 more elements
A molecular complex for enzyme-assisted molecular delivery, the molecular complex comprising:
- a targeting domain;
- a sensor domain, the sensor domain comprising
- a protection segment comprising an RNA portion,
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide
- wherein the sensor domain is bound to the targeting domain through covalent attachment of the targeting domain to the protection segment of the sensor domain at a targeting domain binding portion configured to allow release of the targeting domain upon cleavage of the RNAase H binding site;
- wherein the protection segment, displacement segment, and activation segment are configured so that upon binding of the signal polynucleotide, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding,
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 3′ terminus of the activation segment is adjacent to the 5′ terminus of the targeting domain, and the 5′ terminus of the activation domain is a arranged as a single stranded overhang presented for binding, the 5′ terminus of the toehold segment is adjacent to the 3′ terminus of the protection segment, and the 3′ terminus of the toehold segment is adjacent to the 5′ terminus of the displacement segment.
8 more elements
A molecular complex for enzyme-assisted molecular delivery, the molecular complex comprising:
- a targeting domain;
- a sensor domain, the sensor domain comprising
- a protection segment comprising an RNA portion,
- an activation segment comprising a DNA portion complementary to the RNA portion of the protection segment;
- a displacement segment complementary to the protection segment, the displacement segment complementarily binding the protection segment;
- and
- a toehold segment complementary to a signal polynucleotide
- wherein the sensor domain is bound to the targeting domain through covalent attachment of the targeting domain to the protection segment of the sensor domain at a targeting domain binding portion configured to allow release of the targeting domain upon cleavage of the RNAase H binding site,
- wherein the protection segment, displacement segment, and activation segment are configured so that upon binding of the signal polynucleotide, the displacement segment is displaced from the protection segment and the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding,
- wherein each of the displacement segment, the protection segment, the activation segment and the toehold segment have a 5′ terminus and a 3′ terminus, and
- wherein the 5′ of the activation segment is a single stranded overhang presented for binding, the 3′ terminus of the protection segment is covalently attached to the 5′ terminus of the displacement segment through a polynucleotide linker, and the 3′ terminus of the displacement segment is adjacent to the 5′ terminus of the toehold segment.
8 more elements
Dependencies are inferred from the claim text.