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| Verfasst von: | Medert, Rebekka [VerfasserIn]  |
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| | Thumberger, Thomas [VerfasserIn] |
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| | Tavhelidse-Suck, Tinatini [VerfasserIn] |
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| | Hub, Tobias [VerfasserIn] |
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| | Kellner, Tanja [VerfasserIn] |
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| | Oguchi, Yoko [VerfasserIn] |
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| | Dlugosz, Sascha [VerfasserIn] |
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| | Zimmermann, Frank [VerfasserIn] |
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| | Wittbrodt, Joachim [VerfasserIn] |
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| | Freichel, Marc [VerfasserIn] |
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| Titel: | Efficient single copy integration via homology-directed repair (scHDR) by 5’modification of large DNA donor fragments in mice |
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| Verf.angabe: | Rebekka Medert, Thomas Thumberger, Tinatini Tavhelidse, Tobias Hub, Tanja Kellner, Yoko Oguchi, Sascha Dlugosz, Frank Zimmermann, Joachim Wittbrodt, Marc Freichel |
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| E-Jahr: | 2021 |
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| Jahr: | September 30, 2021 |
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| Umfang: | 37 S. |
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| Illustrationen: | Illustrationen |
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| Fussnoten: | Gesehen am 26.02.2024 |
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| Titel Quelle: | Enthalten in: bioRxiv beta |
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| Ort Quelle: | Cold Spring Harbor : Cold Spring Harbor Laboratory, NY, 2013 |
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| Band/Heft Quelle: | (2021) vom: Sept., Artikel-ID 2021.09.30.462539, Seite 1-37 |
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| Abstract: | CRISPR/Cas approaches have largely replaced conventional gene targeting strategies. However, homology-directed repair (HDR) in the mouse genome is not very efficient, and precisely inserting longer sequences using HDR remains challenging, given that donor constructs preferentially integrate as concatemers. Here, we show that injecting 5’biotinylated donor DNA in mouse embryos at the two-cell stage leads to efficient single-copy HDR (scHDR) alleles. Our dedicated genotyping strategy showed that these alleles occurred with a frequency of 19%, 20%, and 26%, respectively, in three independent gene loci, indicating that scHDR is dramatically boosted by 5’biotinylation. Thus, we suggest that a combination of a 5’biotinylated donor and diligent analysis of concatemer integration are prerequisites for efficiently and reliably generating conditional alleles or other large fragment knock-ins into the mouse genome. |
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| DOI: | doi:10.1101/2021.09.30.462539 |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.1101/2021.09.30.462539 |
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| | Volltext: https://www.biorxiv.org/content/10.1101/2021.09.30.462539v1 |
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| | DOI: https://doi.org/10.1101/2021.09.30.462539 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| K10plus-PPN: | 1881632822 |
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| Verknüpfungen: | → Sammelwerk |
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Efficient single copy integration via homology-directed repair (scHDR) by 5’modification of large DNA donor fragments in mice / Medert, Rebekka [VerfasserIn]; September 30, 2021 (Online-Ressource)
