Highly sensitive detection of microRNAs based on isothermal exponential amplification-assisted generation of catalytic G-quadruplexDNAzyme

Wang, Xin Ping; Yin, Bin; Wang, Ping; Ye, Bang Ce

doi:10.1016/j.bios.2012.10.097

Cited by 79 publications

(24 citation statements)

References 33 publications

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“…Recently, signal amplifying methods based on enzyme reactions have been developed, such as loop-mediated isothermal amplification17, exponential amplification reaction1819, rolling circle amplification (RCA)2021, strand displacement amplification (SDA)2223. SDA and RCA are isothermal DNA amplification techniques based on enzymes.…”

mentioning

confidence: 99%

Label-free detection of microRNA based on coupling multiple isothermal amplification techniques

Zheng

Niu

Wei

et al. 2016

Sci Rep

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MicroRNA (miRNA) was a promising class of cancer biomarkers. Here we developed a label-free method for sensitive measurement of let-7d miRNA based on multiple amplification techniques. The primer will bind to the duplex strand DNA that was formed by stem-loop template and target let-7d to initiate strand displacement amplification (SDA) in tandem. The released single strand DNA will be a primer to bind the circular template to initiate rolling circle amplification (RCA). The products based on multiple amplifications will be detected by a standard fluorimeter with N-methyl mesoporphyrin IX (NMM) as the fluorescent indicator. The proposed method exhibited excellent selectivity and high sensitivity with a detection limit of as low as 1.5 × 10−13 M. Moreover, this methodology was used for the determination of biomolecules in real serum samples with satisfying results.

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mentioning

confidence: 99%

Label-free detection of microRNA based on coupling multiple isothermal amplification techniques

Zheng

Niu

Wei

et al. 2016

Sci Rep

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“…The released substrate triggers the formation of dsDNA concatamers by using SYBR Green I as a signal reporter. In comparison with the previously reported method, this method has the following advantages: 1) high sensitivity to detect a minimum of 10 fM miRNA, 18 2) high selectivity, 19 and 3) no requirement for complex and expensive instrumentation (except for a fluorescence spectrophotometer). The proposed method can be further used to screen the anticancer drugs and might provide a promising approach for the discovery of new anticancer drugs.…”

Section: Resultsmentioning

confidence: 98%

DNAzyme-based probe for circulating microRNA detection in peripheral blood

Shao¹,

Ji²,

Wu³

et al. 2015

DDDT

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BackgroundThe recent discovery of microRNAs (miRNAs) and their extracellular presence suggest a potential role of these regulatory molecules in defining the metastatic potential of cancer cells and mediating the cancer–host communication. This study aims to improve the sensitivity of miRNA detection via DNAzyme-based method and enhance the selectivity by using the DNAzyme-based probe to reduce nonspecific amplification.MethodsThe miRNA probes were chemically synthesized with a phosphate at the 5′ end and purified by polyacrylamide gel electrophoresis. Exosomal RNA from peripheral blood was isolated. Carboxylated magnetic microsphere beads (MBs) were functionalized with streptavidin (SA) according to a previously reported method with some modification. T capture probe-coated SA-MBs (DNA-MBs) were also prepared. The fluorescent spectra were measured using a spectrofluorophotometer.ResultsWe designed an incomplete DNAzyme probe with two stems and one bubble structure as a recognition element for the specific detection of miRNA with high sensitivity. The background effects were decreased with increase of the added of DNA-MBs and capturing times. Therefore, 20 minutes was selected as the optimal concentration in the current study. The fluorescence intensity increases as the hybridization time changed and reached a constant level at 40 minutes, and 1 μM is the optimum signal probe concentration for self-assembled DNA concatemers formation. In the presence of miRNA, the fluorescence of the solution increased with increasing miRNA concentration. There is no obvious fluorescence in the presence of 10 mM of other nontarget DNA.ConclusionA simple, rapid method with high performance has been constructed based on identified circulating miRNA signatures using miRNA-induced DNAzyme. This assay is simple, inexpensive, and sensitive, enabling quantitative detection of as low as 10 fM miRNA.

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“…These exRNAs are loaded onto high-density lipids (HDLs) or bound by AGO2 protein outside the donor cell, thereby protecting miRNA from RNA degrading enzymes. These are known to regulate cell–cell communication and are used as biomarkers in various diseases as these RNAs circulate stably in the blood stream ( 40 , 41 ).…”

Section: Types Of Non-coding Rnasmentioning

confidence: 99%

Export of microRNAs: A Bridge between Breast Carcinoma and Their Neighboring Cells

et al. 2016

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Breast cancer is a leading type of cancer among women in India as well as worldwide. According to the WHO 2015 report, it has been anticipated that there would be a twofold rise in the death due to breast cancer among women. The heterogeneous property of breast carcinoma has been suggested to be linked with dedicated set of communication and signaling pathway with their surroundings, which culminate into progression and development of the cancer. Among the plethora of communication tools in the hand of breast carcinoma cells is the recently appreciated exocytosis of the tightly packed short non-coding RNA molecules, predominantly the microRNAs (miRNAs). Recent studies suggest that miRNAs may work as courier messengers to participate in endocrine and paracrine signaling to facilitate information transfer between breast carcinoma and their neighboring cells. Evidence suggests that breast tumor cells communicate via packaged miRNAs in the tumor-released microvesicles, which enrich the tumor microenvironment. There is a strong view that dissecting out the mechanistic and regulatory aspects of miRNA export and role may uncover many prospects for overcoming the signaling defects and thereby controlling aberrant cell division. The detection of circulating miRNAs associated with breast carcinoma can also be used as biomarkers for early diagnosis. This review article is an attempt to provide updated knowledge on implications of short RNAs and their transport in the breast cancer pathophysiology.

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Highly sensitive detection of microRNAs based on isothermal exponential amplification-assisted generation of catalytic G-quadruplexDNAzyme

Cited by 79 publications

References 33 publications

Label-free detection of microRNA based on coupling multiple isothermal amplification techniques

Label-free detection of microRNA based on coupling multiple isothermal amplification techniques

DNAzyme-based probe for circulating microRNA detection in peripheral blood

Export of microRNAs: A Bridge between Breast Carcinoma and Their Neighboring Cells

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