Side chain modified peptide nucleic acids (PNA) for knock-down of six3 in medaka embryos

doi:10.1186/1472-6750-12-50

BMC Biotechnol. 2012; 12: 50.

Published online 2012 August 17. doi: 10.1186/1472-6750-12-50

PMCID: PMC3469332

Side chain modified peptide nucleic acids (PNA) for knock-down of six3 in medaka embryos

Sebastian Dorn,^#¹ Narges Aghaallaei,^#¹ Gerlinde Jung,^#² Baubak Bajoghli,^1,³ Birgit Werner,⁴ Holger Bock,⁴ Thomas Lindhorst,⁴ and Thomas Czerny^1,²

¹Department for Biomedical Sciences, University of Veterinary Medicine, Veterinärplatz 1, A-1210, Vienna, Austria

²Department for Applied Life Sciences, University of Applied Sciences, FH Campus Wien, Helmut-Qualtinger-Gasse 2, A-1030, Vienna, Austria

³Current address: Director’s Research Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117, Heidelberg, Germany

⁴Ugichem GmbH, Mitterweg 24, A-6020, Innsbruck, Austria

Corresponding author.

^#Contributed equally.

Sebastian Dorn: dornseb/at/hotmail.com; Narges Aghaallaei: aghaallaei/at/immunbio.mpg.de; Gerlinde Jung: gerlinde.jung/at/fh-campuswien.ac.at; Baubak Bajoghli: baubak.bajoghli/at/embl.de; Birgit Werner: birgitwerner/at/ugichem.at; Holger Bock: holgerbock/at/ugichem.at; Thomas Lindhorst: thomaslindhorst/at/ugichem.at; Thomas Czerny: thomas.czerny/at/fh-campuswien.ac.at

Received February 23, 2012; Accepted July 31, 2012.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Synthetic antisense molecules have an enormous potential for therapeutic applications in humans. The major aim of such strategies is to specifically interfere with gene function, thus modulating cellular pathways according to the therapeutic demands. Among the molecules which can block mRNA function in a sequence specific manner are peptide nucleic acids (PNA). They are highly stable and efficiently and selectively interact with RNA. However, some properties of non-modified aminoethyl glycine PNAs (aegPNA) hamper their in vivo applications.

Results

We generated new backbone modifications of PNAs, which exhibit more hydrophilic properties. When we examined the activity and specificity of these novel phosphonic ester PNAs (pePNA) molecules in medaka (Oryzias latipes) embryos, high solubility and selective binding to mRNA was observed. In particular, mixing of the novel components with aegPNA components resulted in mixed PNAs with superior properties. Injection of mixed PNAs directed against the medaka six3 gene, which is important for eye and brain development, resulted in specific six3 phenotypes.

Conclusions

PNAs are well established as powerful antisense molecules. Modification of the backbone with phosphonic ester side chains further improves their properties and allows the efficient knock down of a single gene in fish embryos.

Keywords: PNA, Knock down, Medaka, Six3

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