The human genome is complex, and the functions of many genes and specific genomic elements have not been fully explored. Cultured human cells, from conventional cell lines to iPS cells, are valuable tools that are used to study human genomics in the laboratory; however, the gene targeting efficiencies have generally been low in most of these human cells, making it difficult to assess the functions of these genes. The development of custom-engineered nucleases highlights the possibility of using different approaches to pursue functional genomic studies in cultured human cells. In this chapter, we review the chronological advances made in ZFNs, TALENs and CRISPR/Cas9 systems for gene disruption, locus-specific integration and gene correction. In addition, we summarize the emerging applications of tailor-made DNA binding modules for controlling the transcriptional state, modifying epigenetic marks and for live-cell imaging. The remaining challenges associated with increasing the target specificity, delivery and safety are also discussed.
CITATION STYLE
Li, H. L., & Hotta, A. (2015). Editing cultured human cells: From cell lines to iPS cells. In Targeted Genome Editing Using Site-Specific Nucleases: ZFNs, TALENs, and the CRISPR/Cas9 System (pp. 45–69). Springer Japan. https://doi.org/10.1007/978-4-431-55227-7_3
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