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)

Niss, FridaStockholms universitet,Institutionen för biokemi och biofysik (author)

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.

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Cell- och molekylärbiologi hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Cell and Molecular Biology hsv//eng
NATURVETENSKAP Biologi hsv//swe
NATURAL SCIENCES Biological Sciences hsv//eng
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

Bergqvist, Cecilia,1986-Stockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)cibe6921 (author)
Ström, Anna-Lena,1975-Stockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)astr (author)
Hallberg, EinarStockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)eihal (author)
Stockholms universitetInstitutionen för biokemi och biofysik (creator_code:org_t)

In:The Nuclear Pore ComplexNew York : Humana Press Inc., s. 151-16097810716233679781071623374

By the author/editor: Niss, Frida; Bergqvist, Cecil ...; Ström, Anna-Lena ...; Hallberg, Einar

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Basic Medicine; and Cell and Molecul ...

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