Abstract
Peptide Nucleic Acids (PNAs) are synthetic mimics of natural oligonucleotides, which bind complementary DNA/RNA strands with high sequence specificity. They display numerous advantages, but in vivo applications are still rare. One of the main drawbacks of PNAs application is the poor cellular uptake that could be overcome by using experimental models, in which microinjection techniques allow direct delivery of molecules into eggs. Thus, in this communication, we investigated PNAs efficiency in miR-7 downregulation and compared its effects with those obtained with the commercially available antisense molecule, Antagomir (Dharmacon) in the ascidian Ciona intestinalis. Ascidians are marine invertebrates closely related to vertebrates, in which PNA techniques have not been applied yet. Our results suggested that anti-miR-7 PNAs were able to reach their specific targets in the developing ascidian embryos with high efficiency, as the same effects were obtained with both PNA and Antagomir. To the best of our knowledge, this is the first evidence that unmodified PNAs can be applied in in vivo knockdown strategies when directly injected into eggs.
Highlights
Peptide Nucleic Acids (PNAs) are artificial nucleic acids mimics [1], extensively used for the regulation of gene expression in cellular and molecular systems [2]
In C. intestinalis genome, the miR-7 gene resides within the last intron of the heterogeneous nuclear ribonucleoprotein K gene, oriented in the same direction as the Ci-hnRNP K
Results revealed that miR-7 expression was drastically reduced in embryos injected with with PNA-a7
Summary
Peptide Nucleic Acids (PNAs) are artificial nucleic acids mimics [1], extensively used for the regulation of gene expression in cellular and molecular systems [2]. In PNAs, the neutral pseudo-peptide backbone, based on N-(2-aminoethyl)glycine units (Figure 1A), replaces the negatively charged sugar-phosphate chain of nucleic acids. PNAs can recognize and bind to DNA or RNA sequences according to regular Watson–Crick base pairing rules [3]. Unlike DNA or RNA, PNAs are chemically stable across a wide range of temperatures and pHs, and they are resistant to enzymatic degradation since they are not recognized by nucleases or proteases [4]. The lack of charge repulsion between the neutral PNA strand and the DNA or RNA strand provides extremely stable complexes
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