双链特异性核酸酶的生物学和医学应用
Biological and Medical Applications of Duplex-Specific Nuclease
查看参考文献57篇
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文摘
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双链特异性核酸酶DSN 是一种能高选择性地识别并酶切完全匹配的DNA 双链或者DNA-RNA 杂交双链中的DNA,而对单链DNA 和RNA 几乎没有作用的核酸酶。DSN 酶的上述特点使其在生物和医学等领域广泛应用,主要包括全长cDNA 文库的均一化、单核苷酸多态性(SNP)检测和高通量测序等。近年来, DSN 酶在microRNAs(miRNAs)的检测领域得以长足发展和应用。miRNAs 是一组内源性、非编码短序列 RNAs,在生理和病理过程中具有重要作用,但由于其序列短、丰度低等特点,miRNAs 的检测一直是临床难题。新近研究主要基于DSN 酶信号放大的特点,相继建立了一系列生物传感技术,通过采用不同检测原理如比色法、荧光法、电化学法等实现痕量miRNAs 检测。本文就DSN 酶在miRNAs 检测、SNP 检测、全长 cDNA 文库均一化及高通量测序等方面的生物学应用进行了综述。 |
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其他语种文摘
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Duplex-specific nuclease (DSN) is a type of nuclease,that is isolated from the hepatopancreas of the Kamchatka crab. DSN displays a strong preference for cleaving double-stranded DNA or DNA in DNA-RNA heteroduplexes,and is practically inactive toward single-stranded DNA or RNA. Moreover,this enzyme shows excellent discrimination capability between perfectly and imperfectly matched (up to one mismatch) short duplexes. Owing to its unique feature of cleaving DNA,DSN enzyme is widely applied in the fields of biomedicine and molecular biology,including full-length cDNA library normalization,genomic single-nucleotide polymorphism (SNP) detection and high throughput sequencing. The recent research on DSN are mainly focused on the applications in microRNAs (miRNAs) detection using a DSN-mediated signal amplification strategy. miRNAs are a group of short,endogenous,noncoding RNAs that play vital regulatory roles in physiologic and pathologic processes,including hematopoietic differentiation,cell cycle,regulation,and metabolism. So miRNA is one of the most important biomarkers in individualized treatment,which has great value in terms of improving the diagnosis and treatment of diseases. However,detection of miRNAs is challenging owing to their unique characteristics,including a small size,sequence homology among family members,low abundance in total RNA samples,and susceptibility to degradation in solution. In recent years,isothermal signal amplification and detection of trace miRNA in fluids are reported by many researchers using DSN-mediated biosensors. According to different detection principle of biosensors, DSN-mediated biosensors can be classified as colorimetric,fluorescent,and electrochemical. In this review,we intensively summarize the advantages of DSN in miRNAs detection using DSN-based signal amplification strategy,expanded applications in SNP detection, high throughput sequencing and cDNA library normalization especially when being combined with SMART (Switching Mechanism At 5’end of RNA Transcript) technique. |
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来源
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化学进展
,2014,26(11):1840-1848 【核心库】
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DOI
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10.7536/pc140808
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关键词
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双链特异性核酸酶
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microRNAs检测
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SNP检测
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cDNA文库均一化
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高通量测序
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地址
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第三军医大学附属西南医院输血科, 重庆, 400038
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语种
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中文 |
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文献类型
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综述型 |
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ISSN
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1005-281X |
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学科
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生物化学 |
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基金
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国家自然科学基金项目
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中国军队十二五课题
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重庆市自然科学基金
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第三军医大学基金
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文献收藏号
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CSCD:5274433
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参考文献 共
57
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