Ultrasensitive Detection of Transcription Factors Using Transcription-Mediated Isothermally Exponential Amplification-Induced Chemiluminescence

	Ultrasensitive Detection of Transcription Factors Using Transcription-Mediated Isothermally Exponential Amplification-Induced Chemiluminescence
	Ma, Fei; Yang, Yong; Zhang, Chun-yang
刊名	ANALYTICAL CHEMISTRY
	2014
英文摘要	Transcription factors (TFs) are important cellular components that modulate gene expression, and the malregulation of transcription will lead to a variety ofdiseases such as cancer and developmental syndromes. However, the conventional methods for transcription factor assay are generally cumbersome and costly with low sensitivity. Here, we develop a label-free strategy for ultrasensitive detection of transcription factors using a cascade signal amplification of RNAtranscription, dual isothermally exponential amplification reaction (EXPAR), and G-quadruplex DNAzyme-driven chemiluminescence. Briefly, the specific binding of TF with the detecting probe prevents the cleavage of the detecting probe by exonuclease and subsequently facilitates the conversion of TF signal to abundant RNA triggers in the presence of T7 RNA polymerase. The obtained RNA triggers can initiate the strand displacement amplification to yield abundant DNAzymes and DNA triggers, and the released DNA triggers can further initiate the next rounds of EXPAR reaction. The synergistic operation of dual EXPAR reaction can produce large amounts of DNAzymes, which subsequently catalyze the oxidation of luminol by H2O2 to yield an enhanced chemiluminescencesignal with the assistance of cofactor hemin. Conversely, in the absence of target TF, the naked detecting probes will be completely digested by exonucleases, leading to neither the transcription-mediated EXPAR nor the DNAzyrne-driven chemiluminescence signal. This method has a low detection limitof as low as 6.03 X 10(-15) M and a broad dynamic range from 10 fM to 1 nM and can even measure the NF-kappa B p50 of crude cell nuclear extracts. Moreover, this method can be used to measure a variety of DNA-binding proteins by simply substituting the target-specific binding sequence in the detecting probes.
收录类别	SCI
原文出处	http://pubs.acs.org/doi/abs/10.1021/ac5017369
语种	英语
内容类型	期刊论文
源URL	[http://ir.siat.ac.cn:8080/handle/172644/6135]
专题	深圳先进技术研究院_医药所
作者单位	ANALYTICAL CHEMISTRY
推荐引用方式 GB/T 7714	Ma, Fei,Yang, Yong,Zhang, Chun-yang. Ultrasensitive Detection of Transcription Factors Using Transcription-Mediated Isothermally Exponential Amplification-Induced Chemiluminescence[J]. ANALYTICAL CHEMISTRY,2014.
APA	Ma, Fei,Yang, Yong,&Zhang, Chun-yang.(2014).Ultrasensitive Detection of Transcription Factors Using Transcription-Mediated Isothermally Exponential Amplification-Induced Chemiluminescence.ANALYTICAL CHEMISTRY.
MLA	Ma, Fei,et al."Ultrasensitive Detection of Transcription Factors Using Transcription-Mediated Isothermally Exponential Amplification-Induced Chemiluminescence".ANALYTICAL CHEMISTRY (2014).