title: Genome editing at the crossroads of delivery, specificity, and fidelity.
abstract: Genome editing (GE) entails the modification of specific genomic sequences in living cells for the purpose of determining, changing, or expanding their function(s). Typically, GE occurs after delivering sequence-specific designer nucleases (e.g., ZFNs, TALENs, and CRISPR/Cas9) and donor DNA constructs into target cells. These designer nucleases can generate gene knockouts or gene knock-ins when applied alone or in combination with donor DNA templates, respectively. We review progress in this field, with an emphasis on designer nuclease and donor template delivery into mammalian target cell populations. We also discuss the impact that incremental improvements to these tools are having on the specificity and fidelity attainable with state-of-the-art DNA-editing procedures. Finally, we identify areas that warrant further investigation.
authors.affiliation: Leiden University Medical Center, Department of Molecular Cell Biology, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.
authors.forename: Ignazio
authors.initials: I
authors.lastname: Maggio
authors.affiliation: Leiden University Medical Center, Department of Molecular Cell Biology, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands. Electronic address: m.goncalves@lumc.nl.
authors.forename: Manuel A F V
authors.initials: MA
authors.lastname: Gonçalves
mesh_terms: D000818:Animals; D004247:DNA / Q000737:chemistry; D003851:Deoxyribonucleases / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods / Q000592:standards / Q000639:trends*; D023281:Genomics; D006801:Humans; D008957:Models, Genetic; D015202:Protein Engineering
chemical_list: D004247:DNA; D003851:Deoxyribonucleases
keywords: delivery systems; designer nucleases; donor DNA; gene targeting; genome editing; high fidelity