CRISPR-Cas9介导的人上皮细胞培养中的基因沉默

CRISPR-Cas9-Mediated Gene Silencing in Cultured Human Epithelia.

作者信息

Gago Sara, Overton Nicola L D, Bowyer Paul

机构信息

Manchester Fungal Infection Group, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, Core Technology Facility, The University of Manchester, Manchester, UK.

Clinical Biomarker Centre, CRUK Manchester Institute, The University of Manchester, Manchester, UK.

出版信息

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

DOI:10.1007/978-1-0716-1182-1_4

PMID:33405030

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.

摘要

CRISPR/Cas9技术能够在多种实验系统中实现快速高效的基因组编辑。由于（1）大量现成的商用CRIPSR/Cas9系统，（2）细胞递送效率的提高，以及（3）进行多基因编辑的可能性，使用CRISPR/Cas9进行基因组编辑已成为一种越来越受欢迎的基因工程工具。在此，我们描述了一种在肺支气管上皮细胞中引入单基因破坏的方法。这种方法可用于研究对病原体相互作用重要的宿主因子，或鉴定和研究决定真菌病易感性的遗传标记。