NanoCluster Beacons as reporter probes in rolling circle enhanced enzyme activity detection

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

SN - 2040-3364

VL - 7

SP - 8332

EP - 8337

JO - Nanoscale

JF - Nanoscale

IS - 18

ER -