Marker-free genome editing in Ustilago trichophora with the CRISPR-Cas9 technology

RNA Biology
Simon HuckÜmit Pul

Abstract

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.

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Citations

Nov 6, 2020·Metabolic Engineering·Dongdong ZhaoXueli Zhang
Jul 18, 2019·Computational and Structural Biotechnology Journal·Qiang Wang, Jeffrey J Coleman
Apr 17, 2021·Essays in Biochemistry·Nick WierckxKerstin Schipper
Jun 14, 2021·Fungal Genetics and Biology : FG & B·Chenggang Wang, Jeffrey A Rollins
Oct 21, 2020·Microbiological Research·Rajdeep JaswalT R Sharma

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Methods Mentioned

BETA
genetic modification
gene knock-out
phosphotransferase
PCR

Software Mentioned

GeneArt

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