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Mulholland, C.B.* ; Smets, M.* ; Schmidtmann, E.* ; Leidescher, S.* ; Markaki, Y.* ; Hofweber, M.* ; Qin, W.* ; Manzo, M.* ; Kremmer, E. ; Thanisch, K.* ; Bauer, C.* ; Rombaut, P.* ; Herzog, F.* ; Leonhardt, H.* ; Bultmann, S.*

A modular open platform for systematic functional studies under physiological conditions.

Nucleic Acids Res. 43:e112 (2015)
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Any profound comprehension of gene function requires detailed information about the subcellular localization, molecular interactions and spatio-temporal dynamics of gene products. We developed a multifunctional integrase (MIN) tag for rapid and versatile genome engineering that serves not only as a genetic entry site for the Bxb1 integrase but also as a novel epitope tag for standardized detection and precipitation. For the systematic study of epigenetic factors, including Dnmt1, Dnmt3a, Dnmt3b, Tet1, Tet2, Tet3 and Uhrf1, we generated MIN-tagged embryonic stem cell lines and created a toolbox of prefabricated modules that can be integrated via Bxb1-mediated recombination. We used these functional modules to study protein interactions and their spatio-temporal dynamics as well as gene expression and specific mutations during cellular differentiation and in response to external stimuli. Our genome engineering strategy provides a versatile open platform for efficient generation of multiple isogenic cell lines to study gene function under physiological conditions.
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Publication type Article: Journal article
Document type Scientific Article
Language english
Publication Year 2015
HGF-reported in Year 2015
ISSN (print) / ISBN 0305-1048
e-ISSN 1362-4962
Journal Nucleic Acids Research
Quellenangaben Volume: 43, Issue: 17, Pages: , Article Number: e112 Supplement: ,
Publisher Oxford University Press
Reviewing status Peer reviewed
Institute(s) Institute of Molecular Immunology (IMI)
POF-Topic(s) 30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
Research field(s) Immune Response and Infection
PSP Element(s) G-501793-001
PubMed ID 26007658
DOI 10.1093/nar/gkv550
WOS ID WOS:000366404600006
Scopus ID 84948947590
Erfassungsdatum 2015-05-28
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