DNA recognition and induced genome modification by a hydroxymethyl-γ tail-clamp peptide nucleic acid

• Published in: Cell reports physical science Vol. 4; no. 10; p. 101635
• Main Authors: Oyaghire, Stanley N, Quijano, Elias, Perera, J Dinithi R, Mandl, Hanna K, Saltzman, W Mark, Bahal, Raman, Glazer, Peter M
• Format: Journal Article
• Language: English
• Published: United States 18-10-2023
• ISSN: 2666-3864; 2666-3864
• DOI: 10.1016/j.xcrp.2023.101635

Peptide nucleic acids (PNAs) can target and stimulate recombination reactions in genomic DNA. We have reported that γPNA oligomers possessing the diethylene glycol γ-substituent show improved efficacy over unmodified PNAs in stimulating recombination-induced gene modification. However, this structural modification poses a challenge because of the inherent racemization risk in
-alkylation of the precursory serine side chain. To circumvent this risk and improve γPNA accessibility, we explore the utility of γPNA oligomers possessing the hydroxymethyl-γ moiety for gene-editing applications. We demonstrate that a γPNA oligomer possessing the hydroxymethyl modification, despite weaker preorganization, retains the ability to form a hybrid with the double-stranded DNA target of comparable stability and with higher affinity than that of the diethylene glycol-γPNA. When formulated into poly(lactic-co-glycolic acid) nanoparticles, the hydroxymethyl-γPNA stimulates higher frequencies (≥ 1.5-fold) of gene modification than the diethylene glycol γPNA in mouse bone marrow cells.