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GENOME EDITING

Enhanced precision and efficiency

Nature Biomedical Engineering volume 1, pages 856–857 (2017)Cite this article

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The expression of two DNA repair factors improves the recombination of single-stranded oligodeoxynucleotides with Cas9-induced double-strand breaks, facilitating precise and efficient gene editing.

Interestingly, HDR can also occur with synthetic single-stranded oligodeoxynucleotides (ODNs) as templates4, probably because they mimic natural recombination intermediates. In these cases, ODN sequences are read into the genome in a process called single-stranded template repair (SSTR), which differs from HR and does not depend on RAD51 (Fig. 1). Synthetic ODNs — convenient, comparably inexpensive reagents — can be synthesized with two homology regions of 50–70 nucleotides, up to a limit of ~200 nucleotides. The extent of the practical uses of ODNs in precision gene editing, however, remains unclear because of questions regarding the nature of the recombination mechanism and whether it can be enhanced. Previous efforts for improving this type of editing have focused on optimizing the design of ODNs with regard to the length and symmetry of the homology regions5,6, as well as on modifications to their structure7. Yet, the enzymatic machinery involved remains largely unexplored.

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Fig. 1: Schematic of the DSB-repair pathways used in Cas9-induced genome editing.

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Authors and Affiliations

  1. The Max-Delbrück-Center for Molecular Medicine, Berlin, Germany

    Sanum Bashir & Ralf Kühn

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  1. Sanum Bashir
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  2. Ralf Kühn
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Correspondence to Ralf Kühn.

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Bashir, S., Kühn, R. Enhanced precision and efficiency. Nat Biomed Eng 1, 856–857 (2017). https://doi.org/10.1038/s41551-017-0159-9

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