Abstract
Deep sequencing technology in forward genetics is a powerful tool to identify causal mutations underlying hereditary human diseases. To elucidate the etiological mechanisms, reverse genetics in human cultured cells is useful for generating disease models in vitro. However, the development of reverse genetics has been slow because of the lower efficacy of homologous recombination in almost all mammalian cultured cells. The history of reverse genetics in cultured cells began with the advent of genome editing technology, which could effectively modify the genome via artificial nuclease-induced local DNA repair activity. Bidirectional genetics based on deep sequencing technology and genome editing technology is now an essential approach for clarifying the pathophysiology of hereditary diseases. Here, we provide an overview of the validity of genome editing in cultured cells and its technical problems, discussing the example of centrosome/cilia-related disease models in cultured cells.
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