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Coiled-coil heterodimer-mediated split base editing systems enable flexible and robust nucleotide substitutions.

Shuangshuang Mu1,2,3, Qianru Li1,2,4,5, Menglong Chen6

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Researchers developed a split base editor (BE) system using coiled-coil heterodimers to overcome adeno-associated virus (AAV) packaging limitations. This novel system maintains or improves base editing efficiency for in vivo applications.

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Area of Science:

  • Molecular Biology
  • Gene Editing Technologies

Background:

  • Base editors (BEs) offer precise base substitutions but are too large for adeno-associated virus (AAV) delivery.
  • This size limitation hinders their application in in vivo gene editing therapies.

Purpose of the Study:

  • To engineer a split base editor system compatible with AAV packaging.
  • To assess the editing efficiency and in vivo applicability of the novel split base editor system.

Main Methods:

  • Designed a split base editor system by fusing deaminases to Cas9 nickase via coiled-coil heterodimers.
  • Developed cytidine base editors (CC-CBE) and adenine base editors (CC-ABE) and their derivatives.
  • Evaluated editing efficiency in various cell types and in vivo mouse models.

Main Results:

  • The split base editors (CC-BEs) maintained or enhanced editing efficiency compared to unsplit BEs.
  • CC-CBE achieved up to 12.4-fold enhancement in primary somatic cells.
  • CC-ABE demonstrated successful in vivo A-to-G conversion in Pcsk9 and Dmd genes using dual-AAV vectors in mice.

Conclusions:

  • Developed a simple, universal split base editor strategy to overcome AAV packaging constraints.
  • The CC-BE system enables efficient in vivo base editing without compromising precision.
  • This approach broadens the potential of base editing for therapeutic applications.