Rapid, High-Throughput, and Direct Molecular Beacon Delivery to Human Cancer Cells Using a Nanowire-Incorporated and Pneumatic Pressure-Driven Microdevice.
Tracking and monitoring the intracellular behavior of mRNA is of paramount importance for understanding real-time gene expression in cell biology. To detect specific mRNA sequences, molecular beacons (MBs) have been widely employed as sensing probes. Although numerous strategies for MB delivery into the target cells have been reported, many issues such as the cytotoxicity of the carriers, dependence on the random probability of MB transfer, and critical cellular damage still need to be overcome. Herein, we have developed a nanowire-incorporated and pneumatic pressure-driven microdevice for rapid, high-throughput, and direct MB delivery to human breast cancer MCF-7 cells to monitor survivin mRNA expression. The proposed microdevice is composed of three layers: a pump-associated glass manifold layer, a monolithic polydimethylsiloxane (PDMS) membrane, and a ZnO nanowire-patterned microchannel layer. The MB is immobilized on the ZnO nanowires by disulfide bonding, and the glass manifold and PDMS membrane serve as a microvalve, so that the cellular attachment and detachment on the MB-coated nanowire array can be manipulated. The combination of the nanowire-mediated MB delivery and the microvalve function enable the transfer of MB into the cells in a controllable way with high cell viability and to detect survivin mRNA expression quantitatively after docetaxel treatment.
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Kyung Hoon
KH
Kim
Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Jung
J
Kim
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Jong Seob
JS
Choi
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Sunwoong
S
Bae
Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Donguk
D
Kwon
Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Inkyu
I
Park
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Do Hyun
DH
Kim
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
Tae Seok
TS
Seo
D048430:Cell Shape; D002470:Cell Survival; D005453:Fluorescence; D015972:Gene Expression Regulation, Neoplastic; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000378:metabolism; D061986:MCF-7 Cells; D055616:Microtechnology / Q000295:instrumentation*; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D015335:Molecular Probes / Q000737:chemistry*; D053770:Nanowires / Q000737:chemistry*; D011312:Pressure; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000077022:Survivin; D059008:Time-Lapse Imaging
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D015335:Molecular Probes; D012333:RNA, Messenger; D000077022:Survivin
DNA delivery; cancers; microdevices; molecular beacons; molecular delivery; nanowires; targeted delivery