Side chain modified peptide nucleic acids (PNA) for knock-down of six3 in medaka embryos.
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.
Department for Biomedical Sciences, University of Veterinary Medicine, Veterinärplatz 1, A-1210, Vienna, Austria.
Sebastian
S
Dorn
Narges
N
Aghaallaei
Gerlinde
G
Jung
Baubak
B
Bajoghli
Birgit
B
Werner
Holger
H
Bock
Thomas
T
Lindhorst
Thomas
T
Czerny
D000818:Animals; D001483:Base Sequence; D016373:DNA, Antisense / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics*; D029941:Fish Proteins / Q000235:genetics*; D055786:Gene Knockout Techniques; D018398:Homeodomain Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009990:Oryzias / Q000235:genetics*; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics*; D000097607:Homeobox Protein SIX3
D016373:DNA, Antisense; D005136:Eye Proteins; D029941:Fish Proteins; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D020135:Peptide Nucleic Acids; D000097607:Homeobox Protein SIX3