Peptide Nucleic Acids and Gene Editing: Perspectives on Structure and Repair

Abstract

Unusual nucleic acid structures are salient triggers of endogenous repair and can occur in sequence-specific contexts. Peptide nucleic acids (PNAs) rely on these principles to achieve non-enzymatic gene editing. By forming high-affinity heterotriplex structures within the genome, PNAs have been used to correct multiple human disease-relevant mutations with low off-target effects. Advances in molecular design, chemical modification, and delivery have enabled systemic in vivo application of PNAs resulting in detectable editing in preclinical mouse models. In a model of β-thalassemia, treated animals demonstrated clinically relevant protein restoration and disease phenotype amelioration, suggesting a potential for curative therapeutic application of PNAs to monogenic disorders. This review discusses the rationale and advances of PNA technologies and their application to gene editing with an emphasis on structural biochemistry and repair.

Emerging technologies for cystic fibrosis transmembrane conductance regulator restoration in all people with CF

“…There are several ‐clinical trials exploring the potential of these therapies for treatment of a variety of human diseases including cancer, sickle cell disease and retinal diseases. Many gene editing approaches have been used to correct CFTR mutations including clustered regularly interspaced short palindromic repeats (CRISPR/Cas), 43 zinc finger nucleases (ZFN), 44‐48 transcription activator‐like effector nucleases (TALENS) 49 and triplex forming peptide nucleic acid (PNA)/DNA 50,51 . All the gene editing work in CF is in the preclinical realm 52 .…”

Investigation of the Characteristics of NLS-PNA: Influence of NLS Location on Invasion Efficiency

“…The sequence-specific recognition of double-stranded DNA (dsDNA) has been an important research subject owing to its wide range of potential applications [1][2][3][4][5][6][7]. For this purpose, various methods have been developed, including those that make use of DNA-binding proteins [1,[3][4][5][8][9][10][11][12], small molecules such as minor groove binders [13][14][15][16], and artificial DNA [17][18][19][20][21][22][23][24].…”

“…For this purpose, various methods have been developed, including those that make use of DNA-binding proteins [1,[3][4][5][8][9][10][11][12], small molecules such as minor groove binders [13][14][15][16], and artificial DNA [17][18][19][20][21][22][23][24]. In addition, the recognition of dsDNA by peptide nucleic acid (PNA), an artificial nucleic acid mimic, has been reported by Nielsen et al [7,21,[24][25][26][27][28][29][30][31]. In 1991, PNA was first designed and synthesised as an analogue of natural nucleic acids, where the negatively charged sugar-phosphate backbone of DNA was substituted with an electrostatically neutral artificial N-(2-aminoethyl)glycine backbone ( Figure 1a) [25].…”

“…The thermal stability of the PNA/DNA duplex is enhanced by the formation of stable base pairs between D/U and the natural nucleobases. PNA's unique DNA-recognition mode has been used in many applications, including site-directed mutagenesis [2,7], inhibition of enzymatic activity [26,33,34], and the development of artificial DNA cutters [35], with all of these depending on the sequence specificity of the invasion complex. The pseudopeptide backbone of PNA facilitates easy oligomer synthesis and modification for further improvements and imparts enhanced resistance to enzymatic degradation.…”

Peptide Nucleic Acids: Applications in Biomedical Sciences

“…This property has been used for gene editing in-vitro and in-vivo [ 15 ]. In this Special Issue, Economos et al [ 16 ] review gene editing using a variety of chemically modified PNA (e.g., bisPNA, tail-clamp(tc)PNA, and gamma(γ)PNA) highlighting the clear potential of using this technology to treat monogenic disorders such as β-thalassemia.…”