title: Single-stranded γPNAs 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 (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.
authors.forename: Raman
authors.initials: R
authors.lastname: Bahal
authors.forename: Elias
authors.initials: E
authors.lastname: Quijano
authors.forename: Nicole A
authors.initials: NA
authors.lastname: McNeer
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Liu
authors.forename: Dinesh C
authors.initials: DC
authors.lastname: Bhunia
authors.forename: Francesco
authors.initials: F
authors.lastname: Lopez-Giraldez
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: Fields
authors.forename: William M
authors.initials: WM
authors.lastname: Saltzman
authors.forename: Danith H
authors.initials: DH
authors.lastname: Ly
authors.affiliation: Yale School of Medicine, Dept. of Therapeutic Radiology, P.O. Box 208040, New Haven, Connecticut 06520-8040, USA. peter.glazer@yale.edu.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Glazer
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D018390:Gene Targeting; D049452:Green Fluorescent Proteins / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D019344:Lactic Acid; D051379:Mice; D008822:Mice, Transgenic; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D020135:Peptide Nucleic Acids / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D017393:RNA Editing; D014019:Tissue Donors; D055544:beta-Globins / Q000235:genetics*
chemical_list: D020135:Peptide Nucleic Acids; D055544:beta-Globins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D004247:DNA