ArticlePloS one2018
Efficient genome editing using tRNA promoter-driven CRISPR/Cas9 gRNA in Aspergillus niger.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 65 papers.
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65 citing papers in PubMed.
- Eliminating manganese sensitivity and boosting citric acid production in Aspergillus niger through transporters manipulation.Microbial cell factories · 2026Article
- Establishment of SRLC: a multiplex genome editing technology for Saccharomyces cerevisiae and its application in metabolic engineering of malonyl-CoA pathway.Microbial cell factories · 2026Article
- Characterization of three glycerol transporters with broad sugar specificities from Aspergillus niger.Applied microbiology and biotechnology · 2026Article
- An examination of the quinic acid utilization genes in Aspergillus niger reveals the involvement of 2 pH-dependent permeases.G3 (Bethesda, Md.) · 2025Article
- RNA Polymerase III Promoters Compatible with CRISPR Gene Regulation inACS synthetic biology · 2025Article
- Global survey of secondary metabolism inPNAS nexus · 2025Article
- Comprehensive phenotypic analysis of multiple gene deletions of α-glucan synthase and Crh-transglycosylase gene families inCell surface (Amsterdam, Netherlands) · 2025Article
- Constructing pyrG marker by CRISPR/Cas facilities the highly-efficient precise genome editing on industrial Aspergillus niger strain.Bioprocess and biosystems engineering · 2025Article
- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Metabolic engineering reveals the relative importance of different sugar catabolic pathways during consumption of plant biomass byCurrent research in microbial sciences · 2025Article
- Relative contribution of three transporters to D-xylose uptake inAIMS microbiology · 2025Article
- ReaL-MGE is a tool for enhanced multiplex genome engineering and application to malonyl-CoA anabolism.Nature communications · 2024Article
- Competition between homologous chromosomal DNA and exogenous donor DNA to repair CRISPR/Cas9-induced double-strand breaks in Aspergillus niger.Fungal biology and biotechnology · 2024Article
- NHEJ and HDR can occur simultaneously during gene integration into the genome of Aspergillus niger.Fungal biology and biotechnology · 2024Article
- Increasing the efficiency of CRISPR/Cas9-mediated genome editing in the citrus postharvest pathogen Penicillium digitatum.Fungal biology and biotechnology · 2024Article
- Recent advances in genome-scale engineering inEngineering microbiology · 2024Review
- Utilization of CRISPR-Cas genome editing technology in filamentous fungi: function and advancement potentiality.Frontiers in microbiology · 2024Review
- Establishing a one-step marker-free CRISPR/Cas9 system for industrial Aspergillus niger using counter-selectable marker Ang-ace2.Biotechnology letters · 2023Article
- Genome Mining Reveals a Surprising Number of Sugar Reductases inJournal of fungi (Basel, Switzerland) · 2023Article
- CRISPR/Cas9-Mediated Disruption of theJournal of fungi (Basel, Switzerland) · 2023Article
5 more citing papers are in PubMed but not listed here.
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5 authors.
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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.
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