Title:Single-Stranded γPNAs for In Vivo Site-Specific Genome Editing via Watson-Crick Recognition
Volume: 14
Issue: 5
Author(s): Raman Bahal, Elias Quijano, Nicole A. McNeer, Yanfeng Liu, Dinesh C. Bhunia, Francesco Lopez-Giraldez, Rachel J. Fields, William M. Saltzman, Danith H. Ly and Peter M. Glazer
Affiliation:
Keywords:
β-globin, genome editing, GFP, nanoparticle, PLGA, PNA.
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 (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.
