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

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 az...

Full description

Saved in:
Bibliographic Details
Published inAntimicrobial agents and chemotherapy Vol. 62; no. 9
Main Authors Umeyama, Takashi, Hayashi, Yuta, Shimosaka, Hisaki, Inukai, Tatsuya, Yamagoe, Satoshi, Takatsuka, Shogo, Hoshino, Yasutaka, Nagi, Minoru, Nakamura, Shigeki, Kamei, Katsuhiko, Ogawa, Kenji, Miyazaki, Yoshitsugu
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 01.09.2018
Subjects
Antifungal Agents
Aspergillosis
Aspergillus fumigatus
Cytochrome P-450 Enzyme System
Drug Resistance, Fungal
Fungal Proteins
Gene Editing
Mechanisms of Resistance
Voriconazole
CRISPR
genome editing
Cyp51A
Cas9
azole drugs
antifungal resistance
Online AccessGet full text

Cover

More Information
Summary: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. 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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Citation Umeyama T, Hayashi Y, Shimosaka H, Inukai T, Yamagoe S, Takatsuka S, Hoshino Y, Nagi M, Nakamura S, Kamei K, Ogawa K, Miyazaki Y. 2018. CRISPR/Cas9 genome editing to demonstrate the contribution of Cyp51A Gly138Ser to azole resistance in Aspergillus fumigatus. Antimicrob Agents Chemother 62:e00894-18. https://doi.org/10.1128/AAC.00894-18.
Present address: Hisaki Shimosaka, National Center for Geriatrics and Gerontology, Aichi, Japan.
ISSN:0066-4804
1098-6596
1098-6596
DOI:10.1128/AAC.00894-18