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bioRxiv · 10.1101/348243

Efficient single-copy HDR by 5’ modified long dsDNA donors

Gutierrez-Triana, J. A.·Thumberger, T.·Tavhelidse, T.·Thomas, I.·Wittbrodt, B.·Kellner, T.·Tsingos, E.·Wittbrodt, J.
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Abstract

CRISPR/Cas9 efficiently induces targeted mutations via non-homologous-end-joining but for genome editing, precise, homology-directed repair (HDR) of endogenous DNA stretches is a prerequisite. To favor HDR, many approaches interfere with the repair machinery or manipulate Cas9 itself. Using Medaka we show that the modification of 5 ends of long dsDNA donors strongly enhances HDR, favors efficient single-copy integration by retaining a monomeric donor conformation thus facilitating successful gene replacement or tagging.

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Gutierrez-Triana, J. A., Thumberger, T., Tavhelidse, T., Thomas, I., Wittbrodt, B., Kellner, T., Tsingos, E., Wittbrodt, J.. 2018-06-22. Efficient single-copy HDR by 5’ modified long dsDNA donors. https://doi.org/10.1101/348243

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