Sequence Specificity of Alternating Hydroyprolyl/phosphono Peptide Nucleic Acids against Zebrafish Embryo mRNAs
- 1 July 2004
- journal article
- review article
- Published by Taylor & Francis in Journal of Drug Targeting
- Vol. 12 (6), 363-372
- https://doi.org/10.1080/10611860412331285242
Abstract
Morpholino phosphorodiamidate (MO) DNA mimics display excellent water solubility and hybridization properties toward DNA and RNA, and have been utilized in the model vertebrate zebrafish (Danio rerio) for genome-wide, sequence-based, reverse genetic screens during embryonic development. Peptide nucleic acids (PNAs) exhibit excellent mismatch discrimination, nuclease resistance, and protease resistance, but low solubility. Negatively charged DNA mimics composed of alternating residues of trans-4-hydroxy-L-proline peptide nucleic acid monomers and phosphono peptide nucleic acid monomers (HypNA-pPNA) combine all of the positive features of both MOs and PNAs. Thus, we evaluated PNA oligomers and HypNA-pPNA oligomers as an alternative to MOs for oligonucleotide inhibition of gene expression in zebrafish embryos. We observed that HypNA-pPNA 18-mers displayed comparable potency to MO 25-mers as knockdown agents against chordin, notail and uroD, with greater mismatch stringency. Furthermore, we observed that a specific HypNA-pPNA 18-mer elicited the dharma (bozozok)-/- phenotype in zebrafish embryos, which MO 25-mers do not. These observations validate HypNA-pPNAs as an alternative to MO oligomers for reverse genetic studies. The stronger hybridization and greater specificity of HypNA-pPNAs enable knockdown of mRNAs unaffected by MO oligomers.Keywords
This publication has 19 references indexed in Scilit:
- Phosphono Peptide Nucleic Acids with a Constrained Hydroxyproline-Based BackboneNucleosides, Nucleotides and Nucleic Acids, 2003
- Morpholino Oligos: Making Sense of Antisense?Developmental Biology, 2002
- Morphant technology in model developmental systemsGenesis, 2001
- The homeobox genesvoxandventare redundant repressors of dorsal fates in zebrafishDevelopment, 2001
- PNA-RELATED OLIGONUCLEOTIDE MIMICS AND THEIR EVALUATION FOR NUCLEIC ACID HYBRIDIZATION STUDIES AND ANALYSISNucleosides, Nucleotides and Nucleic Acids, 2001
- Synthesis of polyacrylamides N-substituted with PNA-like oligonucleotide mimics for molecular diagnostic applicationsNucleic Acids Research, 1999
- Phosphonate Analogues of Peptide Nucleic Acids and Related Compounds: Synthesis and Hybridization PropertiesNucleosides and Nucleotides, 1999
- Synthesis and evaluation of some properties of chimeric oligomers containing PNA and phosphono-PNA residuesNucleic Acids Research, 1998
- Peptide Nucleic Acids (PNA): Synthesis, properties and potential applicationsBioorganic & Medicinal Chemistry, 1996
- PNA hybridizes to complementary oligonucleotides obeying the Watson–Crick hydrogen-bonding rulesNature, 1993