Controllable genome editing with split-engineered base editors.

Autor: Berríos KN; Graduate Group in Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Evitt NH; Graduate Group in Cell and Molecular Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., DeWeerd RA; Department of Pediatrics, Washington University School of Medicine, St. Louis, MI, USA., Ren D; Department of Cancer Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Luo M; Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Barka A; Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Wang T; Graduate Group in Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Bartman CR; Epigenetics Institute, Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.; Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Lan Y; Epigenetics Institute, Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA., Green AM; Department of Pediatrics, Washington University School of Medicine, St. Louis, MI, USA., Shi J; Department of Cancer Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jushi@upenn.edu.; Epigenetics Institute, Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jushi@upenn.edu.; Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jushi@upenn.edu., Kohli RM; Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. rkohli@pennmedicine.upenn.edu.; Epigenetics Institute, Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. rkohli@pennmedicine.upenn.edu.; Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. rkohli@pennmedicine.upenn.edu.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2021 Dec; Vol. 17 (12), pp. 1262-1270. Date of Electronic Publication: 2021 Oct 18.
DOI: 10.1038/s41589-021-00880-w
Abstrakt: DNA deaminase enzymes play key roles in immunity and have recently been harnessed for their biotechnological applications. In base editors (BEs), the combination of DNA deaminase mutator activity with CRISPR-Cas localization confers the powerful ability to directly convert one target DNA base into another. While efforts have been made to improve targeting efficiency and precision, all BEs so far use a constitutively active DNA deaminase. The absence of regulatory control over promiscuous deaminase activity remains a major limitation to accessing the widespread potential of BEs. Here, we reveal sites that permit splitting of DNA cytosine deaminases into two inactive fragments, whose reapproximation reconstitutes activity. These findings allow for the development of split-engineered BEs (seBEs), which newly enable small-molecule control over targeted mutator activity. We show that the seBE strategy facilitates robust regulated editing with BE scaffolds containing diverse deaminases, offering a generalizable solution for temporally controlling precision genome editing.
(© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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