{"title":"Thyramide Signal Amplification: New Opportunities for DNA in Situ Hybridization","authors":"E. O. Vorontsova, D. A. Yurchenko, N. V. Shilova","doi":"10.1134/s1990519x23060159","DOIUrl":null,"url":null,"abstract":"<p><b>Abstract</b>—DNA in situ hybridization (ISH) is a valuable technique in molecular cytogenetics for precisely localizing specific DNA sequences on chromosomes. To perform ISH, DNA probes are essential, which can be either commercially available or custom-designed for specific research purposes (homemade probes). However, a drawback of homemade probes is their reduced hybridization signal intensity when they are small. Therefore, it is crucial to develop approaches that optimize the noise/signal ratio when using these DNA probes, which is a current focus in molecular cytogenetics. Tyramide signal amplification (TSA) is a technique that addresses this issue by enabling the visualization of small DNA sequences directly on chromosomes. The TSA system relies on the formation of a covalent bond between electron-rich fragments of sample proteins and tyramide molecules that are linked to a hapten (in chromogenic ISH) or a fluorophore (in fluorescent ISH). This process involves the conversion of tyramide molecules into free radical intermediates by horseradish peroxidase (HRP), followed by the deposition of precipitated molecules in close proximity. As a result, the low-intensity signal is amplified, enhancing the detection sensitivity. TSA serves as an excellent complement to DNA hybridization in situ due to its high sensitivity, allowing the detection of small genomic imbalances. Therefore, it has the potential to be a valuable tool in diagnosing chromosomal rearrangements in clinical practice.</p>","PeriodicalId":9705,"journal":{"name":"Cell and Tissue Biology","volume":"141 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2023-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cell and Tissue Biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1134/s1990519x23060159","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract—DNA in situ hybridization (ISH) is a valuable technique in molecular cytogenetics for precisely localizing specific DNA sequences on chromosomes. To perform ISH, DNA probes are essential, which can be either commercially available or custom-designed for specific research purposes (homemade probes). However, a drawback of homemade probes is their reduced hybridization signal intensity when they are small. Therefore, it is crucial to develop approaches that optimize the noise/signal ratio when using these DNA probes, which is a current focus in molecular cytogenetics. Tyramide signal amplification (TSA) is a technique that addresses this issue by enabling the visualization of small DNA sequences directly on chromosomes. The TSA system relies on the formation of a covalent bond between electron-rich fragments of sample proteins and tyramide molecules that are linked to a hapten (in chromogenic ISH) or a fluorophore (in fluorescent ISH). This process involves the conversion of tyramide molecules into free radical intermediates by horseradish peroxidase (HRP), followed by the deposition of precipitated molecules in close proximity. As a result, the low-intensity signal is amplified, enhancing the detection sensitivity. TSA serves as an excellent complement to DNA hybridization in situ due to its high sensitivity, allowing the detection of small genomic imbalances. Therefore, it has the potential to be a valuable tool in diagnosing chromosomal rearrangements in clinical practice.
摘要--DNA 原位杂交（ISH）是分子细胞遗传学中精确定位染色体上特定 DNA 序列的一项重要技术。要进行原位杂交，DNA 探针是必不可少的，探针可以从市场上购买，也可以为特定研究目的定制（自制探针）。然而，自制探针的一个缺点是，当探针较小时，杂交信号强度会降低。因此，在使用这些 DNA 探针时，开发能优化噪音/信号比的方法至关重要，这也是分子细胞遗传学目前的一个重点。酪胺信号放大（TSA）是一种解决这一问题的技术，它能直接观察染色体上的小 DNA 序列。TSA 系统依赖于样本蛋白质的富电子片段与酪胺分子之间共价键的形成，酪胺分子与合价物（在显色 ISH 中）或荧光团（在荧光 ISH 中）相连。这一过程包括辣根过氧化物酶（HRP）将酪胺分子转化为自由基中间体，然后沉淀的分子在附近沉积。因此，低强度信号被放大，提高了检测灵敏度。TSA 灵敏度高，是 DNA 原位杂交的绝佳补充，可检测微小的基因组失衡。因此，它有望成为临床实践中诊断染色体重排的重要工具。
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The journal publishes papers on vast aspects of cell research, including morphology, biochemistry, biophysics, genetics, molecular biology, immunology. The journal accepts original experimental studies, theoretical articles suggesting novel principles and approaches, presentations of new hypotheses, reviews highlighting major developments in cell biology, discussions. The main objective of the journal is to provide a competent representation and integration of research made on cells (animal and plant cells, both in vivo and in cell culture) offering insight into the structure and functions of live cells as a whole. Characteristically, the journal publishes articles on biology of free-living and parasitic protists, which, unlike Metazoa, are eukaryotic organisms at the cellular level of organization.
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