A highly efficient genome-editing method for Euglena

June 17, 2019

For improvement of industrially useful traits in Euglena

Euglena gracilis is a microalga useful for biofuel and food production. However, the establishment of its effective targeted mutagenesis has been a long-standing challenge, which limited advancement of basic research and breeding in Euglena.

RIKEN CSRS and euglena Co., Ltd. successfully developed an electroporation-based Cas9 RNP delivery method for transgene-free genome editing in E. gracilis. The researchers demonstrated high mutagenesis rates of more than 80% using the developed genome-editing method. Using simultaneous introduction of two Cas9 RNPs targeting different sites, the researchers also successfully generated long deletion mutants of targeted regions in E. gracilis. Moreover, the researchers demonstrated efficient knock-in using Cas9 RNPs with ssODNs in E. gracilis.

These results are expected to greatly contribute to advancing basic research as well as breeding of useful strains in Euglena.

Original article
Plant Biotechnology Journal doi:10.1111/pbi.13174
T. Nomura, K. Inoue, Y. Uehara-Yamaguchi, K. Yamada, O. Iwata, K. Suzuki, K. Mochida,
"Highly efficient transgene-free targeted mutagenesis and single-stranded oligodeoxynucleotidemediated precise knock-in in the industrial microalga Euglena gracilis using Cas9 ribonucleoproteins".
Toshihisa Nomura; Research Scientist
Keiichi Mochida; Team Leader
Bioproductivity Informatics Research Team