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CRISPR/Cas9 Genome Editing To Demonstrate the Contribution of Cyp51A Gly138Ser to Azole Resistance in Aspergillus fumigatus.

ABSTRACT: A pan-azole-resistant Aspergillus fumigatus strain with the cyp51A mutations Gly138Ser and Asn248Lys was isolated from a patient receiving long-term voriconazole treatment. PCR fragments containing cyp51A with the mutations were introduced along with the Cas9 protein and single guide RNA into the azole-resistant/susceptible strains. Recombinant strains showed increased susceptibility via the replacement of Ser138 by glycine. Genetic recombination, which has been hampered thus far in clinical isolates, can now be achieved using CRISPR/Cas9 genome editing.

SUBMITTER: Umeyama T

PROVIDER: S-EPMC6125507 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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CRISPR/Cas9 Genome Editing To Demonstrate the Contribution of Cyp51A Gly138Ser to Azole Resistance in Aspergillus fumigatus.

Umeyama Takashi T Hayashi Yuta Y Shimosaka Hisaki H Inukai Tatsuya T Yamagoe Satoshi S Takatsuka Shogo S Hoshino Yasutaka Y Nagi Minoru M Nakamura Shigeki S Kamei Katsuhiko K Ogawa Kenji K Miyazaki Yoshitsugu Y

Antimicrobial agents and chemotherapy 20180827 9

A pan-azole-resistant Aspergillus fumigatus strain with the cyp51A mutations Gly138Ser and Asn248Lys was isolated from a patient receiving long-term voriconazole treatment. PCR fragments containing cyp51A with the mutations were introduced along with the Cas9 protein and single guide RNA into the azole-resistant/susceptible strains. Recombinant strains showed increased susceptibility via the replacement of Ser138 by glycine. Genetic recombination, which has been hampered thu ...[more]

PMID: 29914956

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