PNA-based artificial nucleases as antisense and anti-miRNA oligonucleotide agents

Abstract

Because of its interesting chemical, physical and biological properties, Peptide Nucleic Acid (PNA) has attracted major attention in molecular biology, for diagnostics purposes and development of biosensors. PNAs have become candidates for gene therapeutic drugs in ANTISENSE (AO) strategy with favorable in vivo biochemical properties. Recently, antisense PNA oligonucleotides have been described in anti-miRNA approach (AMO). We propose PNA-based nucleases as AO and AMO agents. We report the design, synthesis and characterization of two kinds of artificial nucleases composed of a PEG-PNA-PEG domain conjugated to HGG·Cu (A) and DETA (B) as well known cleavage sites. Qualitative (MALDI-TOF) and quantitative (HTS) assays were planned to study nuclease activity of constructs A and B on RNA-3'-FAM target sequence. The results have highlighted the best performance of nuclease B and the relevance of the PEG spacer, in particular for conjugate A, in terms of efficiency of the cleavage, suggesting that conjugates A and B also act as potential antisense and anti-miRNA agents.

Effect of Acylation on the Antimicrobial Activity of Temporin B Analogues

“…For the synthesis of 8 – 12 , we used the peptide KKLLPIVK(Mtt)NLLKSLL anchored to the resin, containing in position 6 a lysine protected with 4‐methyltrityl (Mtt) on the side chain and protected at the N terminus with the Fmoc group. The Mtt group was removed by eight consecutive 1 min treatments of the resin with a solution of trifluoroacetic acid (TFA; 0.1 %)/triisopropylsilane (TIS; 5 %) in CH 2 Cl 2 . The resin was washed with CH 2 Cl 2 and then treated with a solution of DMF/ N , N ‐diisopropylethylamine (DIPEA; 9:1 v / v ) for 5 min.…”

Exploitation of a Very Small Peptide Nucleic Acid as a New Inhibitor of miR-509-3p Involved in the Regulation of Cystic Fibrosis Disease-Gene Expression

“…Furthermore, the cellular uptake behaviour of a PNA is not easily predictable because it is mostly dependent on the PNA base composition and the overall lipophilicity. Recent studies report on the feasibility of a miRNA regulation approach by using unmodified PNAs and PNAs conjugated with peptides or hydrophilic groups [34, 35]. …”

“…In addition, negatively charged PNAs can be obtained by synthesizing PNA-DNA hybrid strands or by attaching negative groups to the PNA monomers [34, 36]. In the last case, cationic lipids can be used as transfection reagents.…”

Versatility of peptide nucleic acids (PNAs): role in chemical biology, drug discovery, and origins of life

“…PNA identifies the difference in species in the translation initiation region of essential genes and selectively inhibits the growth of E. coli , B. subtilis , K. pnuemoniae, and S. enterica serovar Typhimurium in axenic or mixed culture . Some selected most recent publications of PNA in antisense and antigene therapy are summarized in Table ,,…”