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Protein-Functionalized DNA Nanostructures as Tools to Control Transcription in Zebrafish Embryos

Angelin, Alessandro; Kassel, Olivier; Rastegar, Sepand; Strähle, Uwe; Niemeyer, Christof M.

Abstract (englisch):
The unique structure-directing properties of DNA origami nanostructures (DONs) show great potential to specifically manipulate intracellular processes. We report an innovative concept to selectively activate the transcription of a single gene in the developing zebrafish embryo. We reason that engineering a designer transcription factor in which a rigid DON imposes a fixed distance between the DNA-binding domain (DBD) and the transactivation domain (TAD) would allow the selective activation of a gene harboring the same distance between the corresponding transcription factor binding site and the core promoter. As a test case, a rigid tubular DON was designed to separate the DBD of the GAL4 transcription factor and the VP16 viral protein as a TAD. This construct was microinjected in the yolk of one-cell-stage zebrafish embryos, together with a reporter plasmid to assess its functionality. The large DON was efficiently distributed to cells of the developing embryo and showed no signs of toxicity. However, because the DON showed only a cytosolic localization, it did not activate transcription of the reporter gene. Although this work clearly demonstrates that DON microinjection enables the intracellular distribution of multi-protein architectures in most of the cells of the developing zebrafish embryo, further refinements are necessary to enable selective gene activation in vivo.

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Volltext §
DOI: 10.5445/IR/1000065541
Originalveröffentlichung
DOI: 10.1002/open.201600153
Scopus
Zitationen: 4
Web of Science
Zitationen: 3
Coverbild
Zugehörige Institution(en) am KIT Institut für Toxikologie und Genetik (ITG)
Institut für Biologische Grenzflächen (IBG)
Publikationstyp Zeitschriftenaufsatz
Jahr 2017
Sprache Englisch
Identifikator ISSN: 2191-1363, 2191-1355
urn:nbn:de:swb:90-655412
KITopen-ID: 1000065541
HGF-Programm 47.02.01 (POF III, LK 01)
Erschienen in ChemistryOpen
Band 6
Heft 1
Seiten 33–39
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Nachgewiesen in Web of Science
Scopus
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