Marker-free genome editing in Ustilago trichophora with the CRISPR-Cas9 technology
| Autor: | Josephine Bock, Jörg Girardello, Marc Gauert, Simon Huck, Ümit Pul |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Genetics
Gene Editing Genetic Markers 0303 health sciences Base Sequence Ustilago trichophora Cas9 Technical Paper Auxotrophy Cell Biology Biology 03 medical and health sciences 0302 clinical medicine Genome editing 030220 oncology & carcinogenesis CRISPR-Associated Protein 9 Ustilago CRISPR Marker free CRISPR-Cas Systems Chromosomes Fungal Genome Fungal Homologous Recombination Molecular Biology 030304 developmental biology |
| DOI: | 10.6084/m9.figshare.6809195.v1 |
| Popis: | In this communication, we report the adaptation of the CRISPR-Cas9 technology in Ustilago trichophora prototrophic wild-type isolate obtained from its natural host Echinochloa crus-galli. The established CRISPR vector and method enable a rapid and marker-free introduction of Cas9-induced non-homologous end-joining (NHEJ) dependent mutation at the targeted gene. Moreover, the method allows a specific modification of the chromosomal DNA sequence by Cas9-induced homologous recombination using short DNA repair templates. The results demonstrate the applicability of the CRISPR-Cas9 technology in U. trichophora for both gene knock-out by the NHEJ pathway and specific gene modification by templated genome editing, paving the way for rapid metabolic engineering of this Ustilago species for industrial applications. |
| Databáze: | OpenAIRE |
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