García-Rubio María, Barroso Sonia I, Aguilera Andrés
Centro Andaluz de Biología Molecular y Medicina Regenerativa, CABIMER, Universidad de Sevilla, Av. Américo Vespucio 24, 41092, Seville, Spain.
Methods Mol Biol. 2018;1672:347-361. doi: 10.1007/978-1-4939-7306-4_24.
DNA-RNA hybrids form naturally during essential cellular functions such as transcription and replication. However, they may be an important source of genome instability, a hallmark of cancer and genetic diseases. Detection of DNA-RNA hybrids in cells is becoming crucial to understand an increasing number of molecular biology processes in genome dynamics and function and to identify new factors and mechanisms responsible for disease in biomedical research. Here, we describe two different procedures for the reliable detection of DNA-RNA hybrids in the yeast Saccharomyces cerevisiae and in human cells: DNA-RNA Immunoprecipitation (DRIP) and Immunofluorescence.
DNA-RNA杂交体在转录和复制等基本细胞功能过程中自然形成。然而,它们可能是基因组不稳定的一个重要来源,而基因组不稳定是癌症和遗传疾病的一个标志。在细胞中检测DNA-RNA杂交体对于理解基因组动态和功能中越来越多的分子生物学过程,以及在生物医学研究中识别导致疾病的新因素和机制变得至关重要。在这里,我们描述了两种可靠检测酿酒酵母和人类细胞中DNA-RNA杂交体的不同方法:DNA-RNA免疫沉淀(DRIP)和免疫荧光。