Triplex-forming peptide nucleic acids (PNAs) facilitate gene editing by stimulating recombination of donor
DNAs within genomic DNA via site-specific formation of altered helical structures that further stimulate DNA repair.
However, PNAs designed for triplex formation are sequence restricted to homopurine sites. Herein we describe a novel
strategy where next generation single-stranded gamma PNAs (γPNAs) containing miniPEG substitutions at the gamma
position can target genomic DNA in mouse bone marrow at mixed-sequence sites to induce targeted gene editing. In addition
to enhanced binding, γPNAs confer increased solubility and improved formulation into poly(lactic-co-glycolic acid)
(PLGA) nanoparticles for efficient intracellular delivery. Single-stranded γPNAs induce targeted gene editing at frequencies
of 0.8% in mouse bone marrow cells treated ex vivo and 0.1% in vivo via IV injection, without detectable toxicity.
These results suggest that γPNAs may provide a new tool for induced gene editing based on Watson-Crick recognition
without sequence restriction.
Keywords: β-globin, genome editing, GFP, nanoparticle, PLGA, PNA.
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