Using cells encapsulated in agarose microbeads to analyse nuclear structure and functions

    Research output: Chapter in Book/Report/Conference proceedingChapter

    Abstract

    It is now generally agreed that the nuclei of higher eukaryotes, and particularly of mammalian cells, are highly structured and that different aspects of this structure contribute to the regulation of function (1, 2). Despite the general consensus, the key mechanisms that link nuclear structure and function have proved elusive. A major reason for this is a lack of techniques that allow nuclei to be manipulated in a way that preserves the complex architectural features that are present in vivo. Historically, significant progress in understanding the makeup of nuclei from mammalian cells has been made using cells that are permeabilised in a physiological buffer after being encapsulated in agarose microbeads. By using such beads, cells are protected from shear forces that otherwise can degrade crucial elements of the architecture that it is essential to preserve. © 2009 Humana Press.
    Original languageEnglish
    Title of host publicationMethods in Molecular Biology|Methods Mol. Biol.
    PublisherHumana Press, Inc
    Pages139-158
    Number of pages19
    Volume464
    DOIs
    Publication statusPublished - 2008

    Keywords

    • Agarose-encapsulated cells
    • Cell extraction
    • Chromatin
    • DNA loops
    • DNA replication
    • Nucleoskeleton
    • RNA transcription

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