Department of Clinical Chemistry, Laboratory Genetic Metabolic Diseases, Amsterdam UMC - University of Amsterdam, AZ, Amsterdam, the Netherlands.
Methods Mol Biol. 2023;2643:233-245. doi: 10.1007/978-1-0716-3048-8_17.
The development and application of the CRISPR-Cas9 technology for genome editing of mammalian cells have opened up a wealth of possibilities for genetically modifying and manipulating human cells, and use in functional studies or therapeutic approaches.Here we describe the approach that we have been using successfully to generate multiple human cell lines with targeted (partial) gene deletions, i.e., knockout cells, or human cells with modified genomic nucleotide sequences, i.e., knock-in cells, in genes encoding known or putative proteins involved in peroxisome biogenesis or peroxisomal functions.
CRISPR-Cas9 技术在哺乳动物细胞基因组编辑中的发展和应用为遗传修饰和操作人类细胞开辟了丰富的可能性,并可用于功能研究或治疗方法。在这里,我们描述了一种成功用于生成具有靶向(部分)基因缺失的多种人类细胞系的方法,即敲除细胞,或具有修饰的基因组核苷酸序列的人类细胞,即敲入细胞,这些基因编码已知或假定参与过氧化物酶体生物发生或过氧化物酶体功能的蛋白质。