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A heterodimer of evolved designer-recombinases precisely excises a human genomic DNA locus.


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.

SUBMITTER: Lansing F 

PROVIDER: S-EPMC7107906 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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A heterodimer of evolved designer-recombinases precisely excises a human genomic DNA locus.

Lansing Felix F   Paszkowski-Rogacz Maciej M   Schmitt Lukas Theo LT   Schneider Paul Martin PM   Rojo Romanos Teresa T   Sonntag Jan J   Buchholz Frank F  

Nucleic acids research 20200101 1


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 exp  ...[more]

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