A heterodimer of evolved designer-recombinases precisely excises a human genomic DNA locus.
Nucleic Acids Res
; 48(1): 472-485, 2020 01 10.
Article
em En
| MEDLINE
| ID: mdl-31745551
ABSTRACT
Site-specific recombinases (SSRs) such as the Cre/loxP system are useful genome engineering tools that can be repurposed by altering their DNA-binding specificity. However, SSRs that delete a natural sequence from the human genome have not been reported thus far. Here, we describe the generation of an SSR system that precisely excises a 1.4 kb fragment from the human genome. Through a streamlined process of substrate-linked directed evolution we generated two separate recombinases that, when expressed together, act as a heterodimer to delete a human genomic sequence from chromosome 7. Our data indicates that designer-recombinases can be generated in a manageable timeframe for precision genome editing. A large-scale bioinformatics analysis suggests that around 13% of all human protein-coding genes could be targetable by dual designer-recombinase induced genomic deletion (dDRiGD). We propose that heterospecific designer-recombinases, which work independently of the host DNA repair machinery, represent an efficient and safe alternative to nuclease-based genome editing technologies.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Cromossomos Humanos Par 7
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Sequência de Bases
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Genoma Humano
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Deleção de Sequência
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DNA Nucleotidiltransferases
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Edição de Genes
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article