Graphene-oxide quenching-based molecular beacon imaging of exosome-mediated transfer of neurogenic miR-193a on microfluidic platform

Hyun Jeong Oh; Jaehoon Kim; Hyejin Park; Seok Chung; Do Won Hwang; Dong Soo Lee

doi:10.1016/j.bios.2018.11.027

Graphene-oxide quenching-based molecular beacon imaging of exosome-mediated transfer of neurogenic miR-193a on microfluidic platform

Hyun Jeong Oh, Jaehoon Kim, Hyejin Park, Seok Chung, Do Won Hwang, Dong Soo Lee

Research output: Contribution to journal › Article › peer-review

39 Citations (Scopus)

Abstract

Graphene-oxide (GO) quenching-based molecular beacon was developed for rapid and sensitive detection of RNAs in living cells and tissues. Here, we applied GO quenching-based molecular beacon sensor to visualize neurogenic miR-193a levels delivered via exosomes during cell-non-autonomous neurogenesis in neural progenitor cells on a microfluidic platform. Exosomal transport was visualized using CD63-RFP plasmid vector, and FAM-labeled peptide nucleic acid (PNA) probe for the miR-193 sequence was designed to detect endogenous miR-193 expression. Fluorescence signals of FAM-PNA193a-GO were recovered in dibutyryl-cAMP-induced F11 cells, resulting from increased expression of miR-193a after neuronal differentiation. We observed delivery of miR-193a-containing exosomes released from differentiated donor F11 cells to recipient undifferentiated F11 cells. Fluorescence recovery was evident in exosome-stimulated recipient individual F11 cells in the microfluidic system. We propose molecular beacon imaging using PNA-GO complex for visualization of individual cellular expression of mature microRNAs. This system reveals the precise spatial localization and temporal sequences of mature miRNAs by intercellular exosomal delivery of messages for processes such as cell-non-autonomous neurogenesis.

Original language	English
Pages (from-to)	647-656
Number of pages	10
Journal	Biosensors and Bioelectronics
Volume	126
DOIs	https://doi.org/10.1016/j.bios.2018.11.027
Publication status	Published - 2019 Feb 1

Bibliographical note

Funding Information:
This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2017R1A6A3A01007423 ), and Radiation Technology R&D program through the National Research Foundation of Korea founded by the Ministry of Science and ICT ( NRF-2017M2A2A7A02019899 ), and by the National Research Foundation of Korea ( NRF-2017R1A2B3007701 ) grant funded by the Korea government(MEST), and by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea ( HI14C3344 and HI14C1277 ), and National Research Foundation of Korea grant funded by the Korea government (MSIP) ( 2015M3C7A1028926 and 2017M3C7A1048079 ).

Publisher Copyright:
© 2018 Elsevier B.V.

Keywords

Exosome-mediated neurogenesis
Graphene oxide (GO)
Microfluidic platform
Molecular beacon sensor
Peptide nucleic acid (PNA)
miRNA-193a

ASJC Scopus subject areas

Biotechnology
Biophysics
Biomedical Engineering
Electrochemistry

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10.1016/j.bios.2018.11.027

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@article{d2172a3bc11348e99919332e2cd45698,

title = "Graphene-oxide quenching-based molecular beacon imaging of exosome-mediated transfer of neurogenic miR-193a on microfluidic platform",

abstract = "Graphene-oxide (GO) quenching-based molecular beacon was developed for rapid and sensitive detection of RNAs in living cells and tissues. Here, we applied GO quenching-based molecular beacon sensor to visualize neurogenic miR-193a levels delivered via exosomes during cell-non-autonomous neurogenesis in neural progenitor cells on a microfluidic platform. Exosomal transport was visualized using CD63-RFP plasmid vector, and FAM-labeled peptide nucleic acid (PNA) probe for the miR-193 sequence was designed to detect endogenous miR-193 expression. Fluorescence signals of FAM-PNA193a-GO were recovered in dibutyryl-cAMP-induced F11 cells, resulting from increased expression of miR-193a after neuronal differentiation. We observed delivery of miR-193a-containing exosomes released from differentiated donor F11 cells to recipient undifferentiated F11 cells. Fluorescence recovery was evident in exosome-stimulated recipient individual F11 cells in the microfluidic system. We propose molecular beacon imaging using PNA-GO complex for visualization of individual cellular expression of mature microRNAs. This system reveals the precise spatial localization and temporal sequences of mature miRNAs by intercellular exosomal delivery of messages for processes such as cell-non-autonomous neurogenesis.",

keywords = "Exosome-mediated neurogenesis, Graphene oxide (GO), Microfluidic platform, Molecular beacon sensor, Peptide nucleic acid (PNA), miRNA-193a",

author = "Oh, {Hyun Jeong} and Jaehoon Kim and Hyejin Park and Seok Chung and Hwang, {Do Won} and Lee, {Dong Soo}",

note = "Funding Information: This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2017R1A6A3A01007423 ), and Radiation Technology R&D program through the National Research Foundation of Korea founded by the Ministry of Science and ICT ( NRF-2017M2A2A7A02019899 ), and by the National Research Foundation of Korea ( NRF-2017R1A2B3007701 ) grant funded by the Korea government(MEST), and by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea ( HI14C3344 and HI14C1277 ), and National Research Foundation of Korea grant funded by the Korea government (MSIP) ( 2015M3C7A1028926 and 2017M3C7A1048079 ). Publisher Copyright: {\textcopyright} 2018 Elsevier B.V.",

year = "2019",

month = feb,

day = "1",

doi = "10.1016/j.bios.2018.11.027",

language = "English",

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AU - Oh, Hyun Jeong

AU - Kim, Jaehoon

AU - Park, Hyejin

AU - Chung, Seok

AU - Hwang, Do Won

AU - Lee, Dong Soo

N1 - Funding Information: This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education ( NRF-2017R1A6A3A01007423 ), and Radiation Technology R&D program through the National Research Foundation of Korea founded by the Ministry of Science and ICT ( NRF-2017M2A2A7A02019899 ), and by the National Research Foundation of Korea ( NRF-2017R1A2B3007701 ) grant funded by the Korea government(MEST), and by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea ( HI14C3344 and HI14C1277 ), and National Research Foundation of Korea grant funded by the Korea government (MSIP) ( 2015M3C7A1028926 and 2017M3C7A1048079 ). Publisher Copyright: © 2018 Elsevier B.V.

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N2 - Graphene-oxide (GO) quenching-based molecular beacon was developed for rapid and sensitive detection of RNAs in living cells and tissues. Here, we applied GO quenching-based molecular beacon sensor to visualize neurogenic miR-193a levels delivered via exosomes during cell-non-autonomous neurogenesis in neural progenitor cells on a microfluidic platform. Exosomal transport was visualized using CD63-RFP plasmid vector, and FAM-labeled peptide nucleic acid (PNA) probe for the miR-193 sequence was designed to detect endogenous miR-193 expression. Fluorescence signals of FAM-PNA193a-GO were recovered in dibutyryl-cAMP-induced F11 cells, resulting from increased expression of miR-193a after neuronal differentiation. We observed delivery of miR-193a-containing exosomes released from differentiated donor F11 cells to recipient undifferentiated F11 cells. Fluorescence recovery was evident in exosome-stimulated recipient individual F11 cells in the microfluidic system. We propose molecular beacon imaging using PNA-GO complex for visualization of individual cellular expression of mature microRNAs. This system reveals the precise spatial localization and temporal sequences of mature miRNAs by intercellular exosomal delivery of messages for processes such as cell-non-autonomous neurogenesis.

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KW - Graphene oxide (GO)

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KW - Molecular beacon sensor

KW - Peptide nucleic acid (PNA)

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ER -

Graphene-oxide quenching-based molecular beacon imaging of exosome-mediated transfer of neurogenic miR-193a on microfluidic platform

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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