CRISPR-Cas9-Mediated Gene Silencing in Cultured Human Epithelia

Methods Mol Biol. 2021:2260:37-47. doi: 10.1007/978-1-0716-1182-1_4.

Abstract

CRISPR/Cas9 technology enables rapid and efficient genome editing in a variety of experimental systems. Genome editing using CRISPR/Cas9 has become an increasingly popular genetic engineering tool due to (1) an extensive array of commercial ready-to-use CRIPSR/Cas9 systems, (2) improved efficiency of cell delivery, and (3) the possibility to do multigene editing. Here, we describe a method to introduce single gene disruption in lung bronchial epithelial cells. This approach can be used to study host factors important for pathogen interaction or to identify and study genetic markers determining susceptibility to fungal disease.

Keywords: CRISPR/Cas9; Genome editing; Host–fungi interaction.

MeSH terms

  • Bronchi / metabolism*
  • Bronchi / microbiology
  • CRISPR-Associated Protein 9 / genetics*
  • CRISPR-Associated Protein 9 / metabolism
  • CRISPR-Cas Systems*
  • Cell Line
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Epithelial Cells / metabolism*
  • Epithelial Cells / microbiology
  • Fungi / pathogenicity*
  • Gene Editing*
  • Gene Expression Regulation
  • Gene Silencing*
  • Host-Parasite Interactions
  • Humans
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • RNA, Guide, CRISPR-Cas Systems / metabolism

Substances

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