Single-stranded ?PNAs for in vivo site-specific genome editing via Watson-Crick recognition.

ABSTRACT: 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 (?PNAs) 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, ?PNAs confer increased solubility and improved formulation into poly(lactic-co-glycolic acid) (PLGA) nanoparticles for efficient intracellular delivery. Single-stranded ?PNAs 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 ?PNAs may provide a new tool for induced gene editing based on Watson-Crick recognition without sequence restriction.

SUBMITTER: Bahal R 

PROVIDER: S-EPMC4333085 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature