A DNAzyme-mediated target-initiated rolling circle amplification strategy based on microchip platform for the detection of apurinic/apyrimidinic endonuclease 1 at single-cell level

2021 ◽  
Author(s):  
Chunhuan Xu ◽  
Shengyu Chen ◽  
Jingjin Zhao ◽  
Xiaoshu Luo ◽  
Shulin Zhao
Keyword(s):  
Single Cell ◽  
Signal Amplification ◽  
Rolling Circle Amplification ◽  
Single Cell Level ◽  
Cell Level ◽  
Rolling Circle ◽  
Amplification Strategy

A DNAzyme-mediated target-initiated rolling circle signal amplification strategy based on microchip platform was developed for detecting apurinic/apyrimidine endonuclease 1 (APE1) at the single-cell level. This strategy was applied for assay...

scholarly journals Quantitative Determination of Free-DNA Uptake in River Bacteria at the Single-Cell Level by In Situ Rolling-Circle Amplification

2006 ◽  
Vol 72 (9) ◽  
pp. 6248-6256 ◽  
Author(s):  
Fumito Maruyama ◽  
Katsuji Tani ◽  
Takehiko Kenzaka ◽  
Nobuyasu Yamaguchi ◽  
Masao Nasu
Keyword(s):  
Single Cell ◽  
Gene Amplification ◽  
Rolling Circle Amplification ◽  
Extracellular Dna ◽  
Single Cell Level ◽  
Dna Uptake ◽  
Cell Level ◽  
Indigenous Bacteria ◽  
Rolling Circle

ABSTRACT Detection of plasmid DNA uptake in river bacteria at the single-cell level was carried out by rolling-circle amplification (RCA). Uptake of a plasmid containing the green fluorescent protein gene (gfp) by indigenous bacteria from two rivers in Osaka, Japan, was monitored for 506 h using this in situ gene amplification technique with optimized cell permeabilization conditions. Plasmid uptake determined by in situ RCA was compared to direct counts of cells expressing gfp under fluorescence microscopy to examine differences in detection sensitivities between the two methods. Detection of DNA uptake as monitored by in situ RCA was 20 times higher at maximum than that by direct counting of gfp-expressing cells. In situ RCA could detect bacteria taking up the plasmid in several samples in which no gfp-expressing cells were apparent, indicating that in situ gene amplification techniques can be used to determine accurate rates of extracellular DNA uptake by indigenous bacteria in aquatic environments.

scholarly journals MiRNA Detection Using a Rolling Circle Amplification and RNA-Cutting Allosteric Deoxyribozyme Dual Signal Amplification Strategy

Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 222
Author(s):  
Chenxin Fang ◽  
Ping Ouyang ◽  
Yuxing Yang ◽  
Yang Qing ◽  
Jialun Han ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Rolling Circle Amplification ◽  
Padlock Probe ◽  
Rolling Circle ◽  
Sensing Platform ◽  
Mirna Detection ◽  
Substrate Cleavage ◽  
Amplification Strategy ◽  
Circular Template ◽  
Dual Signal Amplification

A microRNA (miRNA) detection platform composed of a rolling circle amplification (RCA) system and an allosteric deoxyribozyme system is proposed, which can detect miRNA-21 rapidly and efficiently. Padlock probe hybridization with the target miRNA is achieved through complementary base pairing and the padlock probe forms a closed circular template under the action of ligase; this circular template results in RCA. In the presence of DNA polymerase, RCA proceeds and a long chain with numerous repeating units is formed. In the presence of single-stranded DNA (H1 and H2), multi-component nucleic acid enzymes (MNAzymes) are formed that have the ability to cleave substrates. Finally, substrates containing fluorescent and quenching groups and magnesium ions are added to the system to activate the MNAzyme and the substrate cleavage reaction, thus achieving fluorescence intensity amplification. The RCA–MNAzyme system has dual signal amplification and presents a sensing platform that demonstrates broad prospects in the analysis and detection of nucleic acids.

Label-Free Telomerase Detection in Single Cell Using a Five-Base Telomerase Product-Triggered Exponential Rolling Circle Amplification Strategy

ACS Sensors ◽  
2019 ◽  
Vol 4 (4) ◽  
pp. 1090-1096 ◽  
Author(s):  
XiaoYu Li ◽  
YunXi Cui ◽  
YiChen Du ◽  
AnNa Tang ◽  
DeMing Kong
Keyword(s):  
Single Cell ◽  
Rolling Circle Amplification ◽  
Label Free ◽  
Rolling Circle ◽  
Amplification Strategy

A triple-amplification strategy for sensitive detection of telomerase at the single-cell level

2018 ◽  
Vol 54 (67) ◽  
pp. 9317-9320 ◽  
Author(s):  
Chen-chen Li ◽  
Yan Zhang ◽  
Wen-jing Liu ◽  
Chun-yang Zhang
Keyword(s):  
Single Cell ◽  
Cancer Cells ◽  
Sensitive Detection ◽  
Single Cell Level ◽  
Cell Level ◽  
Amplification Strategy

We develop a triple-amplification strategy for sensitive detection of telomerase from cancer cells at the single-cell level.

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