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Homologous recombination-mediated gene targeting is a powerful tool to study genes’ functions and has been instrumental for breakthroughs in mice genetics and disease models. However, such technology has not been broadly applied in regular research labs due to the low targeting efficiency in human cell lines and demand of expertise. The newly discovered transcription activator-like effector nuclease (TALEN) technology appears to be effective, less laborious and involves minimal expertise. In this study, we designed and constructed several TALEN constructs for nuclear genome targeting. We also modified TALE nucleases to make them more suitable for mitochondria genome editing. We showed that a homozygous gene knockout cell line can be obtained in one round of gene targeting in one month (compared to the 4-6 months required for traditional homologous recombination based studies). Mitochondrial genome can be readily edited as well, suggesting a large application of TALEN technology in mitochondria research. Our data indicates that TALEN technology has strong potential for routine gene knockout in regular research labs.
Last Modified December 14, 2012