TY - JOUR
T1 - Ultrasensitive FRET-based DNA sensor using PNA/DNA hybridization
AU - Yang, Lan Hee
AU - Ahn, Dong June
AU - Koo, Eunhae
N1 - Funding Information:
This study was supported by a grant from the Fundamental R&D Program for Core Technology of Materials funded (Grant 10047831 ) by the Ministry of Trade, Industry & Energy (MOTIE), Republic of Korea.
Publisher Copyright:
© 2016
PY - 2016/12/1
Y1 - 2016/12/1
N2 - In the diagnosis of genetic diseases, rapid and highly sensitive DNA detection is crucial. Therefore, many strategies for detecting target DNA have been developed, including electrical, optical, and mechanical methods. Herein, a highly sensitive FRET based sensor was developed by using PNA (Peptide Nucleic Acid) probe and QD, in which red color QDs are hybridized with capture probes, reporter probes and target DNAs by EDC-NHS coupling. The hybridized probe with target DNA gives off fluorescent signal due to the energy transfer from QD to Cy5 dye in the reporter probe. Compared to the conventional DNA sensor using DNA probes, the DNA sensor using PNA probes shows higher FRET factor and efficiency due to the higher reactivity between PNA and target DNA. In addition, to elicit the effect of the distance between the donor and the acceptor, we have investigated two types of the reporter probes having Cy5 dyes attached at the different positions of the reporter probes. Results show that the shorter the distance between QDs and Cy5s, the stronger the signal intensity. Furthermore, based on the fluorescence microscopy images using microcapillary chips, the FRET signal is enhanced to be up to 276% times stronger than the signal obtained using the cuvette by the fluorescence spectrometer. These results suggest that the PNA probe system conjugated with QDs can be used as ultrasensitive DNA nanosensors.
AB - In the diagnosis of genetic diseases, rapid and highly sensitive DNA detection is crucial. Therefore, many strategies for detecting target DNA have been developed, including electrical, optical, and mechanical methods. Herein, a highly sensitive FRET based sensor was developed by using PNA (Peptide Nucleic Acid) probe and QD, in which red color QDs are hybridized with capture probes, reporter probes and target DNAs by EDC-NHS coupling. The hybridized probe with target DNA gives off fluorescent signal due to the energy transfer from QD to Cy5 dye in the reporter probe. Compared to the conventional DNA sensor using DNA probes, the DNA sensor using PNA probes shows higher FRET factor and efficiency due to the higher reactivity between PNA and target DNA. In addition, to elicit the effect of the distance between the donor and the acceptor, we have investigated two types of the reporter probes having Cy5 dyes attached at the different positions of the reporter probes. Results show that the shorter the distance between QDs and Cy5s, the stronger the signal intensity. Furthermore, based on the fluorescence microscopy images using microcapillary chips, the FRET signal is enhanced to be up to 276% times stronger than the signal obtained using the cuvette by the fluorescence spectrometer. These results suggest that the PNA probe system conjugated with QDs can be used as ultrasensitive DNA nanosensors.
KW - DNA sensor
KW - Fret
KW - Microcapillary
KW - PNA
KW - QD
UR - http://www.scopus.com/inward/record.url?scp=84978288410&partnerID=8YFLogxK
U2 - 10.1016/j.msec.2016.07.021
DO - 10.1016/j.msec.2016.07.021
M3 - Article
C2 - 27612755
AN - SCOPUS:84978288410
SN - 0928-4931
VL - 69
SP - 625
EP - 630
JO - Materials Science and Engineering C
JF - Materials Science and Engineering C
ER -