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The effects of external Mn2+ concentration on hyphal morphology and citric acid production are mediated primarily by the NRAMP-family transporter DmtA in Aspergillus niger.

Author(s): Fejes B, Ouedraogo JP, Fekete E, Sándor E, Flipphi M, Soós Á, Molnár ÁP, Kovács B, Kubicek CP, Tsang A, Karaffa L

Microb Cell Fact. 2020 Jan 30;19(1):17 Authors: Fejes B, Ouedraogo JP, Fekete E, Sándor E, Flipphi M, Soós Á, Molnár ÁP, Kovács B, Kubicek CP, Tsang A, Karaffa L

Article GUID: 32000778
A set of isogenic auxotrophic strains for constructing multiple gene deletion mutants and parasexual crossings in Aspergillus niger.

Author(s): Niu J, Arentshorst M, Seelinger F, Ram AF, Ouedraogo JP

Arch Microbiol. 2016 Nov;198(9):861-8 Authors: Niu J, Arentshorst M, Seelinger F, Ram AF, Ouedraogo JP

Article GUID: 27251039
Efficient genome editing using tRNA promoter-driven CRISPR/Cas9 gRNA in Aspergillus niger.

Author(s): Song L, Ouedraogo JP, Kolbusz M, Nguyen TTM, Tsang A

PLoS One. 2018;13(8):e0202868 Authors: Song L, Ouedraogo JP, Kolbusz M, Nguyen TTM, Tsang A

Article GUID: 30142205
Title:Efficient genome editing using tRNA promoter-driven CRISPR/Cas9 gRNA in Aspergillus niger.
Authors:Song L,  Ouedraogo JP,  Kolbusz M,  Nguyen TTM,  Tsang A
Link:https://www.ncbi.nlm.nih.gov/pubmed/30142205?dopt=Abstract
DOI:10.1371/journal.pone.0202868
Category:PLoS One
PMID:30142205
Dept Affiliation: GENOMICS
1 Centre for Structural and Functional Genomics, Concordia University, Montreal, Canada.

Description:

Efficient genome editing using tRNA promoter-driven CRISPR/Cas9 gRNA in Aspergillus niger.

PLoS One. 2018;13(8):e0202868

Authors: Song L, Ouedraogo JP, Kolbusz M, Nguyen TTM, Tsang A

Abstract

As a powerful tool for fast and precise genome editing, the CRISPR/Cas9 system has been applied in filamentous fungi to improve the efficiency of genome alteration. However, the method of delivering guide RNA (gRNA) remains a bottleneck in performing CRISPR mutagenesis in Aspergillus species. Here we report a gRNA transcription driven by endogenous tRNA promoters which include a tRNA gene plus 100 base pairs of upstream sequence. Co-transformation of a cas9-expressing plasmid with a linear DNA coding for gRNA demonstrated that 36 of the 37 tRNA promoters tested were able to generate the intended mutation in A. niger. When gRNA and cas9 were expressed in a single extra-chromosomal plasmid, the efficiency of gene mutation was as high as 97%. Co-transformation with DNA template for homologous recombination, the CRISPR/Cas9 system resulted ~42% efficiency of gene replacement in a strain with a functioning non-homologous end joining machinery (kusA+), and an efficiency of >90% gene replacement in a kusA- background. Our results demonstrate that tRNA promoter-mediated gRNA expressions are reliable and efficient in genome editing in A. niger.

PMID: 30142205 [PubMed - indexed for MEDLINE]