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Programmable and multiparameter DNA-based logic platform for cancer recognition and targeted therapy.

You M, Zhu G, Chen T, Donovan MJ, Tan W - J. Am. Chem. Soc. (2014)

Bottom Line: The specific inventory of molecules on diseased cell surfaces (e.g., cancer cells) provides clinicians an opportunity for accurate diagnosis and intervention.With the discovery of panels of cancer markers, carrying out analyses of multiple cell-surface markers is conceivable.The success of this strategy demonstrates the potential of DNA nanotechnology in facilitating targeted disease diagnosis and effective therapy.

View Article: PubMed Central - PubMed

Affiliation: Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center for Molecular Engineering and Theranostics, Hunan University , Changsha, Hunan 410082, China.

ABSTRACT
The specific inventory of molecules on diseased cell surfaces (e.g., cancer cells) provides clinicians an opportunity for accurate diagnosis and intervention. With the discovery of panels of cancer markers, carrying out analyses of multiple cell-surface markers is conceivable. As a trial to accomplish this, we have recently designed a DNA-based device that is capable of performing autonomous logic-based analysis of two or three cancer cell-surface markers. Combining the specific target-recognition properties of DNA aptamers with toehold-mediated strand displacement reactions, multicellular marker-based cancer analysis can be realized based on modular AND, OR, and NOT Boolean logic gates. Specifically, we report here a general approach for assembling these modular logic gates to execute programmable and higher-order profiling of multiple coexisting cell-surface markers, including several found on cancer cells, with the capacity to report a diagnostic signal and/or deliver targeted photodynamic therapy. The success of this strategy demonstrates the potential of DNA nanotechnology in facilitating targeted disease diagnosis and effective therapy.

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Construction of three-inputcell-surface logic gates. (A) Symbols,truth tables, and experimental schemes of three-input “a ANDb NOT c” gate. (B) The realization of several three-input cell-surfacelogic gates, using rationally designed tagged-Sgc4f/Sgc8c/TE17 aptamerpairs and CEM cells as examples (the experimental schemes for eachgate are shown in the SI). Cell viabilitytest was performed after visible irradiation for 3 h and subsequentgrowth for 48 h (*: p-value <0.05; **: p-value <0.001; by comparison with each irradiated celltype only, n = 3). Bracketed letter-labeled strand(e.g., [c] strand) is complementary to the strand labeled with thesame letter (e.g., c strand).
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fig3: Construction of three-inputcell-surface logic gates. (A) Symbols,truth tables, and experimental schemes of three-input “a ANDb NOT c” gate. (B) The realization of several three-input cell-surfacelogic gates, using rationally designed tagged-Sgc4f/Sgc8c/TE17 aptamerpairs and CEM cells as examples (the experimental schemes for eachgate are shown in the SI). Cell viabilitytest was performed after visible irradiation for 3 h and subsequentgrowth for 48 h (*: p-value <0.05; **: p-value <0.001; by comparison with each irradiated celltype only, n = 3). Bracketed letter-labeled strand(e.g., [c] strand) is complementary to the strand labeled with thesame letter (e.g., c strand).

Mentions: In a practical application of such higher-order logicoperation, after a group of specific oligonucleotide tag-connectedaptamers bound with cell membrane markers, three types of actuatorstrands will be added to realize the identification and targeted therapy.The first is a reporter strand, the fluorophore- or therapeutic reagent-labeledoligonucleotide that provides the fluorescence readout for detectionor therapeutic functions after binding with the specific tags on thecellular membrane. The second is one or more gate strands, which isprehybridized with the reporter strand with a leftover toehold region.Several gate strands can function in cascade when they are designedto be partially complementary with each other (Figure 3A). The gate strands function by hybridizing with membranetags of the complementary sequence and through strand displacementreactions, free the reporter strand for final signaling. The thirdis an assistant strand, whose sequence is complementary to one ofthe gate strands and can displace/free the reporter strand. However,the presence of a cell membrane tag with the same sequence as thegate strand will inhibit the activation process, since the assistantstrand will preferentially bind to fully complementary (36 bp) freetag, instead of the short toehold region (8 bp) of the gate strand.28 The assistant strand functions for the realizationof a NOT gate, i.e., the presence of a cell membrane input inhibitsthe activation of the fluorescence signal or therapeutic effect. Thesethree types of strands are rationally designed in our platform tofunction together as the barcode reader and actuator.


Programmable and multiparameter DNA-based logic platform for cancer recognition and targeted therapy.

You M, Zhu G, Chen T, Donovan MJ, Tan W - J. Am. Chem. Soc. (2014)

Construction of three-inputcell-surface logic gates. (A) Symbols,truth tables, and experimental schemes of three-input “a ANDb NOT c” gate. (B) The realization of several three-input cell-surfacelogic gates, using rationally designed tagged-Sgc4f/Sgc8c/TE17 aptamerpairs and CEM cells as examples (the experimental schemes for eachgate are shown in the SI). Cell viabilitytest was performed after visible irradiation for 3 h and subsequentgrowth for 48 h (*: p-value <0.05; **: p-value <0.001; by comparison with each irradiated celltype only, n = 3). Bracketed letter-labeled strand(e.g., [c] strand) is complementary to the strand labeled with thesame letter (e.g., c strand).
© Copyright Policy - editor-choice
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4308741&req=5

fig3: Construction of three-inputcell-surface logic gates. (A) Symbols,truth tables, and experimental schemes of three-input “a ANDb NOT c” gate. (B) The realization of several three-input cell-surfacelogic gates, using rationally designed tagged-Sgc4f/Sgc8c/TE17 aptamerpairs and CEM cells as examples (the experimental schemes for eachgate are shown in the SI). Cell viabilitytest was performed after visible irradiation for 3 h and subsequentgrowth for 48 h (*: p-value <0.05; **: p-value <0.001; by comparison with each irradiated celltype only, n = 3). Bracketed letter-labeled strand(e.g., [c] strand) is complementary to the strand labeled with thesame letter (e.g., c strand).
Mentions: In a practical application of such higher-order logicoperation, after a group of specific oligonucleotide tag-connectedaptamers bound with cell membrane markers, three types of actuatorstrands will be added to realize the identification and targeted therapy.The first is a reporter strand, the fluorophore- or therapeutic reagent-labeledoligonucleotide that provides the fluorescence readout for detectionor therapeutic functions after binding with the specific tags on thecellular membrane. The second is one or more gate strands, which isprehybridized with the reporter strand with a leftover toehold region.Several gate strands can function in cascade when they are designedto be partially complementary with each other (Figure 3A). The gate strands function by hybridizing with membranetags of the complementary sequence and through strand displacementreactions, free the reporter strand for final signaling. The thirdis an assistant strand, whose sequence is complementary to one ofthe gate strands and can displace/free the reporter strand. However,the presence of a cell membrane tag with the same sequence as thegate strand will inhibit the activation process, since the assistantstrand will preferentially bind to fully complementary (36 bp) freetag, instead of the short toehold region (8 bp) of the gate strand.28 The assistant strand functions for the realizationof a NOT gate, i.e., the presence of a cell membrane input inhibitsthe activation of the fluorescence signal or therapeutic effect. Thesethree types of strands are rationally designed in our platform tofunction together as the barcode reader and actuator.

Bottom Line: The specific inventory of molecules on diseased cell surfaces (e.g., cancer cells) provides clinicians an opportunity for accurate diagnosis and intervention.With the discovery of panels of cancer markers, carrying out analyses of multiple cell-surface markers is conceivable.The success of this strategy demonstrates the potential of DNA nanotechnology in facilitating targeted disease diagnosis and effective therapy.

View Article: PubMed Central - PubMed

Affiliation: Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center for Molecular Engineering and Theranostics, Hunan University , Changsha, Hunan 410082, China.

ABSTRACT
The specific inventory of molecules on diseased cell surfaces (e.g., cancer cells) provides clinicians an opportunity for accurate diagnosis and intervention. With the discovery of panels of cancer markers, carrying out analyses of multiple cell-surface markers is conceivable. As a trial to accomplish this, we have recently designed a DNA-based device that is capable of performing autonomous logic-based analysis of two or three cancer cell-surface markers. Combining the specific target-recognition properties of DNA aptamers with toehold-mediated strand displacement reactions, multicellular marker-based cancer analysis can be realized based on modular AND, OR, and NOT Boolean logic gates. Specifically, we report here a general approach for assembling these modular logic gates to execute programmable and higher-order profiling of multiple coexisting cell-surface markers, including several found on cancer cells, with the capacity to report a diagnostic signal and/or deliver targeted photodynamic therapy. The success of this strategy demonstrates the potential of DNA nanotechnology in facilitating targeted disease diagnosis and effective therapy.

Show MeSH
Related in: MedlinePlus