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Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery

Rees, Holly A ; Komor, Alexis C ; Yeh, Wei-Hsi ; Caetano-Lopes, Joana ; Warman, Matthew ; Edge, Albert S B ; Liu, David R

Nature communications, 2017-06, Vol.8 (1), p.15790-15790, Article 15790 [Periódico revisado por pares]

England: Nature Publishing Group

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Título:
Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery
Autor: Rees, Holly A ; Komor, Alexis C ; Yeh, Wei-Hsi ; Caetano-Lopes, Joana ; Warman, Matthew ; Edge, Albert S B ; Liu, David R
Assuntos: Animals ; Cell Line ; Chromatography ; CRISPR-Cas Systems ; DNA - genetics ; DNA - metabolism ; Editors ; Gene Editing ; Gene loci ; Genomes ; Laboratories ; Mice ; Mutation ; Otology ; Plasmids ; Protein Engineering ; Proteins ; Ribonucleoproteins - genetics ; Ribonucleoproteins - metabolism ; Zebrafish
É parte de: Nature communications, 2017-06, Vol.8 (1), p.15790-15790, Article 15790
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Descrição: We recently developed base editing, a genome-editing approach that enables the programmable conversion of one base pair into another without double-stranded DNA cleavage, excess stochastic insertions and deletions, or dependence on homology-directed repair. The application of base editing is limited by off-target activity and reliance on intracellular DNA delivery. Here we describe two advances that address these limitations. First, we greatly reduce off-target base editing by installing mutations into our third-generation base editor (BE3) to generate a high-fidelity base editor (HF-BE3). Next, we purify and deliver BE3 and HF-BE3 as ribonucleoprotein (RNP) complexes into mammalian cells, establishing DNA-free base editing. RNP delivery of BE3 confers higher specificity even than plasmid transfection of HF-BE3, while maintaining comparable on-target editing levels. Finally, we apply these advances to deliver BE3 RNPs into both zebrafish embryos and the inner ear of live mice to achieve specific, DNA-free base editing in vivo.
Editor: England: Nature Publishing Group
Idioma: Inglês

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Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery

Rees, Holly A ; Komor, Alexis C ; Yeh, Wei-Hsi ; Caetano-Lopes, Joana ; Warman, Matthew ; Edge, Albert S B ; Liu, David R

England: Nature Publishing Group

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