Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review
NanoCluster Beacons as reporter probes in rolling circle enhanced enzyme activity detection. / Juul, Sissel; Obliosca, Judy M; Liu, Cong; Liu, Yen-Liang; Chen, Yu-An; Imphean, Darren M; Knudsen, Birgitta R; Ho, Yi-Ping; Leong, Kam W; Yeh, Hsin-Chih.
In: Nanoscale, Vol. 7, No. 18, 30.04.2015, p. 8332-8337.Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review
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TY - JOUR
T1 - NanoCluster Beacons as reporter probes in rolling circle enhanced enzyme activity detection
AU - Juul, Sissel
AU - Obliosca, Judy M
AU - Liu, Cong
AU - Liu, Yen-Liang
AU - Chen, Yu-An
AU - Imphean, Darren M
AU - Knudsen, Birgitta R
AU - Ho, Yi-Ping
AU - Leong, Kam W
AU - Yeh, Hsin-Chih
PY - 2015/4/30
Y1 - 2015/4/30
N2 - As a newly developed assay for the detection of endogenous enzyme activity at the single-catalytic-event level, Rolling Circle Enhanced Enzyme Activity Detection (REEAD) has been used to measure enzyme activity in both single human cells and malaria-causing parasites, Plasmodium sp. Current REEAD assays rely on organic dye-tagged linear DNA probes to report the rolling circle amplification products (RCPs), the cost of which may hinder the widespread use of REEAD. Here we show that a new class of activatable probes, NanoCluster Beacons (NCBs), can simplify the REEAD assays. Easily prepared without any need for purification and capable of large fluorescence enhancement upon hybridization, NCBs are cost-effective and sensitive. Compared to conventional fluorescent probes, NCBs are also more photostable. As demonstrated in reporting the human topoisomerases I (hTopI) cleavage-ligation reaction, the proposed NCBs suggest a read-out format attractive for future REEAD-based diagnostics.
AB - As a newly developed assay for the detection of endogenous enzyme activity at the single-catalytic-event level, Rolling Circle Enhanced Enzyme Activity Detection (REEAD) has been used to measure enzyme activity in both single human cells and malaria-causing parasites, Plasmodium sp. Current REEAD assays rely on organic dye-tagged linear DNA probes to report the rolling circle amplification products (RCPs), the cost of which may hinder the widespread use of REEAD. Here we show that a new class of activatable probes, NanoCluster Beacons (NCBs), can simplify the REEAD assays. Easily prepared without any need for purification and capable of large fluorescence enhancement upon hybridization, NCBs are cost-effective and sensitive. Compared to conventional fluorescent probes, NCBs are also more photostable. As demonstrated in reporting the human topoisomerases I (hTopI) cleavage-ligation reaction, the proposed NCBs suggest a read-out format attractive for future REEAD-based diagnostics.
U2 - 10.1039/c5nr01705j
DO - 10.1039/c5nr01705j
M3 - Journal article
C2 - 25901841
VL - 7
SP - 8332
EP - 8337
JO - Nanoscale
JF - Nanoscale
SN - 2040-3364
IS - 18
ER -