Zusammenfassung
DNA-Nukleasen, darunter Zink-Finger-Nukleasen (ZFN), Transkriptionsaktivator-ähnliche Endonukleasen (TALENS) und Meganukleasen, besitzen lange Erkennungsstellen und Schneidedomänen und sind daher in der Lage, DNA auf sehr spezifische Weise zu schneiden. Diese molekularen Scheren bewirken gezielte genetische Veränderungen, indem sie die DNA-Mutationsrate durch Induktion von Doppelstrangbrüchen an einer vorbestimmten genomischen Stelle erhöhen. Im Vergleich zum konventionellen Gen-Targeting auf der Basis homologer Rekombination können DNA-Nukleasen die Targeting-Rate um das bis zu 10.000-fache erhöhen, und die Genunterbrechung durch mutagene DNA-Reparatur wird mit einer ähnlichen Häufigkeit stimuliert. Die erfolgreiche Anwendung verschiedener DNA-Nukleasen wurde bei einer Vielzahl von Organismen nachgewiesen, darunter Insekten, Amphibien, Pflanzen, Fadenwürmer und Säugetiere, einschließlich Nutztiere. Kürzlich wurde eine weitere neue Klasse von molekularen Scheren beschrieben, die kurze RNA-Sequenzen verwenden, um eine bestimmte Stelle im Genom zu verändern (Abb. 7.1). CRISPR/CAS9 geht auf einen bakteriellen Abwehrmechanismus zurück und kann so programmiert werden, dass es auf fast jede Stelle im Genom verändern kann. Die einfache und kostengünstige Möglichkeit, mit Hilfe von DNA-Nukleasen sehr spezifische genetische Veränderungen zu erzeugen, hat die Produktion von gentechnisch veränderten Nutztieren revolutioniert. Aktuelle Ergebnisse deuten darauf hin, dass DNA-Nukleasen erfolgreich in einem breiten Spektrum von Organismen eingesetzt werden können, was sie für ein besseres Verständnis komplexer physiologischer Systeme, für die Erzeugung genetisch veränderter Tiere, einschließlich der Schaffung von Großtiermodellen für menschliche Krankheiten und für die Schaffung spezifischer Zelllinien nützlich macht. Gentechnische Veränderungen könnten auch den Tierschutz verbessern, indem sie das Enthornen oder die Geschlechtsbestimmung überflüssig oder Nutztiere resistent/resistent gegen bestimmte Krankheitserreger machen. Nutztiere mit einem gewünschten Phänotyp oder Merkmal können mit bisher unbekannter Präzision und Leichtigkeit und innerhalb eines sehr kurzen Zeitraums erzeugt werden, was vor Einführung der DNA Nukleasen als unmöglich galt. Dieses Kapitel bietet einen aktuellen Überblick über DNA-Nukleasen und die ihnen zugrunde liegenden Mechanismen und konzentriert sich auf ihre Verwendung in der Tierproduktion. Dabei ist zu bedenken, dass zum Zeitpunkt der Abfassung dieses Kapitels noch keine gentechnisch veränderten Nutztiere in die Nahrungskette gelangt sind oder für die Herstellung von aus Nutztieren gewonnenen Produkten verwendet wurden.
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Petersen, B. (2023). DNA-Nukleasen und ihre Verwendung in der Viehzucht. In: Niemann, H., Wrenzycki, C. (eds) Biotechnologie bei Nutztieren 2. Springer, Cham. https://doi.org/10.1007/978-3-031-26042-1_7
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