Highly efficient single base editing in Aspergillus niger with CRISPR/Cas9 cytidine deaminase fusion.

Abstract:

:Classic genome editing tools including ZFN, TALEN, and CRISPR/Cas9 rely on DNA double-strand breaks for genome editing. To prevent the potential hazard caused by double-strand breaks (DSBs), a series of single base editing tools that convert cytidine (C) to thymine (T) without DSBs have been developed extensively in multiple species. Herein, we report for the first time that C was converted to T with a high frequency in the filamentous fungi Aspergillus niger by fusing cytidine deaminase and Cas9 nickase. Using the CRISPR/Cas9-dependent base editor and inducing nonsense mutations via single base editing, we inactivated the uridine auxotroph gene pyrG and the pigment gene fwnA with an efficiency of 47.36%-100% in A.niger. At the same time, the single-base editing results of the non-phenotypic gene prtT showed an efficiency of 60%. The editable window reached 8 bases (from C2 to C9 in the protospacer) in A. niger. Overall, we successfully constructed a single base editing system in A. niger. This system provides a more convenient tool for investigating gene function in A. niger, and provides a new tool for genetic modification in filamentous fungi.

journal_name

Microbiol Res

journal_title

Microbiological research

authors

Huang L,Dong H,Zheng J,Wang B,Pan L

doi

10.1016/j.micres.2019.03.007

subject

Has Abstract

pub_date

2019-01-01 00:00:00

pages

44-50

eissn

0944-5013

issn

1618-0623

pii

S0944-5013(18)31379-X

journal_volume

223-225

pub_type

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