CRISPR/Cas9 genome editing technology in filamentous fungi: progress and perspective

Appl Microbiol Biotechnol. 2019 Sep;103(17):6919-6932. doi: 10.1007/s00253-019-10007-w. Epub 2019 Jul 22.

Abstract

Filamentous fungi play an important role in human health and industrial/agricultural production. With the increasing number of full genomes available for fungal species, the study of filamentous fungi has brought about a wider range of genetic manipulation opportunities. However, the utilization of traditional methods to study fungi is time consuming and laborious. Recent rapid progress and wide application of a versatile genome editing technology, i.e., the CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR-related nuclease 9) system, has revolutionized biological research and has many innovative applications in a wide range of fields showing great promise in research and application of filamentous fungi. In this review, we introduce the CRISPR/Cas9 genome editing technology focusing on its application in research of filamentous fungi and we discuss the general considerations of genome editing using CRISPR/Cas9 system illustrating vector construction, multiple editing strategies, technical consideration of different sizes of homology arms on genome editing efficiency, off-target effects, and different transformation methodologies. In addition, we discuss the challenges encountered using CRISPR/Cas9 technology and give the perspectives of future applications of CRISPR/Cas9 technology for basic research and practical application of filamentous fungi.

Keywords: CRISPR/Cas9; Filamentous fungi; Genome editing; Off-target.

Publication types

  • Review

MeSH terms

  • CRISPR-Associated Protein 9 / genetics
  • CRISPR-Cas Systems* / genetics
  • Fungi / genetics*
  • Gene Editing*
  • Gene Targeting
  • Genome, Fungal / genetics*
  • Industrial Microbiology
  • Mutation
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Transformation, Genetic

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • CRISPR-Associated Protein 9