Capítulos (Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER))

URI permanente para esta colecciónhttps://hdl.handle.net/11441/11091

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    Acceso AbiertoCapítulo de Libro
    Analysis of repair of replication-born double-strand breaks by sister chromatid recombination in yeast
    (Elsevier, 2021) Gómez González, Belén; Ortega Moreno, Pedro; Aguilera López, Andrés; Universidad de Sevilla. Departamento de Genética
    The repair of DNA double-strand breaks is crucial for cell viability and the maintenance of genome integrity. When present, the intact sister chromatid is used as the preferred repair template to restore the genetic information by homologous recombination. Although the study of the factors involved in sister chromatid recombination is hampered by the fact that both sister chromatids are indistinguishable, genetic and molecular systems based on DNA repeats have been developed to overcome this problem. In particular, the use of site-specific nucleases capable of inducing DNA nicks that replication converts into double-strand breaks has enabled the specific study of the repair of such replication-born double strand breaks by sister chromatid recombination. In this chapter, we describe detailed protocols for determining the efficiency and kinetics of this recombination reaction as well as for the genetic quantification of recombination products.
  • Miniatura
    Acceso AbiertoCapítulo de Libro
    Detection of DNA-RNA hybrids in vivo
    (Humana Press Inc., 2018) García Rubio, María Luisa; Barroso Ceballos, Sonia Inés; Aguilera López, Andrés; Universidad de Sevilla. Departamento de Genética
    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.