Monitoring of Chromatin Organization at the Nuclear Pore Complex, Inner Nuclear Membrane, and Nuclear Interior in Live Cells by Fluorescence Ratiometric Imaging of Chromatin (FRIC)
2022 (English)In: The Nuclear Pore Complex: Methods and Protocols / [ed] Martin W. Goldberg, New York: Humana Press Inc. , 2022, p. 151-160Chapter in book (Refereed)
Abstract [en]
The image analysis tool FRIC (Fluorescence Ratiometric Imaging of Chromatin) quantitatively monitors dynamic spatiotemporal distribution of euchromatin and total chromatin in live cells. A vector (pTandemH) assures stoichiometrically constant expression of the histone variants Histone 3.3 and Histone 2B, fused to EGFP and mCherry, respectively. Quantitative ratiometric (H3.3/H2B) imaging displayed a concentrated distribution of heterochromatin in the periphery of U2OS cell nuclei. As a proof of concept, peripheral heterochromatin responded to experimental manipulation of histone acetylation as well as expression of the mutant lamin A protein “progerin,” which causes Hutchinson-Gilford Progeria Syndrome. In summary FRIC is versatile, unbiased, robust, requires a minimum of experimental steps and is suitable for screening purposes.
Place, publisher, year, edition, pages
New York: Humana Press Inc. , 2022. p. 151-160
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 2502
Keywords [en]
Chromatin, Fluorescence ratiometric, Live imaging, Nuclear membrane, Nuclear pore complex, histone, lamin A, cell nucleus, cell nucleus membrane, fluorescence, genetics, HeLa cell line, heterochromatin, human, metabolism, nuclear pore, HeLa Cells, Histones, Humans, Lamin Type A, Nuclear Envelope
National Category
Cell and Molecular Biology Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-206355DOI: 10.1007/978-1-0716-2337-4_10PubMedID: 35412237Scopus ID: 2-s2.0-85128080535ISBN: 978-1-0716-2336-7 (print)ISBN: 978-1-0716-2337-4 (electronic)OAI: oai:DiVA.org:su-206355DiVA, id: diva2:1669218
2022-06-142022-06-142022-09-23Bibliographically approved