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The function of tcf3 in medaka embryos: efficient knockdown with pePNAs

Doenz, Gerlinde ; Dorn, Sebastian ; Aghaallaei, Narges ; Bajoghli, Baubak ; Riegel, Elisabeth ; Aigner, Michaela ; Bock, Holger ; Werner, Birgit ; Lindhorst, Thomas ; Czerny, Thomas

In: BMC Biotechnology, 18 (2018), Nr. 1. pp. 2-14. ISSN 1472-6750

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Download (1MB) | Lizenz: Creative Commons LizenzvertragThe function of tcf3 in medaka embryos: efficient knockdown with pePNAs by Doenz, Gerlinde ; Dorn, Sebastian ; Aghaallaei, Narges ; Bajoghli, Baubak ; Riegel, Elisabeth ; Aigner, Michaela ; Bock, Holger ; Werner, Birgit ; Lindhorst, Thomas ; Czerny, Thomas underlies the terms of Creative Commons Attribution 4.0

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Abstract

Background: The application of antisense molecules, such as morpholino oligonucleotides, is an efficient method of gene inactivation in vivo. We recently introduced phosphonic ester modified peptide nucleic acids (PNA) for in vivo loss-of-function experiments in medaka embryos. Here we tested novel modifications of the PNA backbone to knockdown the medaka tcf3 gene.

Results: A single tcf3 gene exists in the medaka genome and its inactivation strongly affected eye development of the embryos, leading to size reduction and anophthalmia in severe cases. The function of Tcf3 strongly depends on co-repressor interactions. We found interactions with Groucho/Tle proteins to be most important for eye development. Using a dominant negative approach for combined inactivation of all groucho/tle genes also resulted in eye phenotypes, as did interference with three individual tle genes.

Conclusions: Our results show that side chain modified PNAs come close to the knockdown efficiency of morpholino oligonucleotides in vivo. A single medaka tcf3 gene combines the function of the two zebrafish paralogs hdl and tcf3b. In combination with Groucho/Tle corepressor proteins Tcf3 acts in anterior development and is critical for eye formation.

Item Type: Article
Journal or Publication Title: BMC Biotechnology
Volume: 18
Number: 1
Publisher: BioMed Central
Place of Publication: London
Date Deposited: 23 Apr 2018 11:37
Date: 2018
ISSN: 1472-6750
Page Range: pp. 2-14
Faculties / Institutes: Service facilities > Centre for Organismal Studies Heidelberg (COS)
Subjects: 570 Life sciences
610 Medical sciences Medicine
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