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Controllable genome editing with split-engineered base editors.

Kiara N Berríos1, Niklaus H Evitt2, Rachel A DeWeerd3

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|October 19, 2021
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Summary
This summary is machine-generated.

Researchers developed split-engineered base editors (seBEs) to control DNA deaminase activity. This breakthrough enables precise, temporally regulated genome editing using small molecules.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Editing

Background:

  • DNA deaminase enzymes are crucial for immunity and biotechnological applications.
  • Base editors (BEs) combine DNA deaminases with CRISPR-Cas for targeted DNA base conversion.
  • Current BEs lack regulatory control over deaminase activity, limiting their potential.

Purpose of the Study:

  • To develop a method for controlling the activity of DNA deaminases within base editors.
  • To engineer split-engineered base editors (seBEs) for regulated genome editing.
  • To enable small-molecule control over targeted mutator activity in precision genome editing.

Main Methods:

  • Identified sites for splitting DNA cytosine deaminases into two inactive fragments.
  • Reconstituted deaminase activity upon reapproximation of the split fragments.
  • Developed split-engineered BEs (seBEs) incorporating this split-enzyme strategy.

Main Results:

  • Demonstrated that split DNA cytosine deaminases can be reconstituted to regain activity.
  • Successfully developed seBEs that allow for small-molecule-mediated control of deaminase activity.
  • Showcased robust, regulated editing using seBEs with diverse deaminase scaffolds.

Conclusions:

  • The split-enzyme strategy offers a generalizable solution for temporal control of precision genome editing.
  • seBEs provide a novel platform for regulating targeted mutator activity.
  • This approach overcomes the limitation of constitutively active deaminases in current base editors.