On February 24, 2013, "Nature Methods" published a research paper "TALEN mediated precise genome modification by homologous recombination in zebrafish" mainly in the laboratory of Professor Zhang Bo. The research team used TALE nuclease (TALEN) to cause double-stranded DNA breaks in the zebrafish tyrosine hydroxylase (th) gene, while using classic long double-stranded DNA as a target carrier, successfully passed the same Source recombination integrates the GFP gene into the endogenous th gene, and proves that this precisely modified mutant allele can be stably inherited into future generations.
Using homologous recombination to modify the genome precisely is very important for studying gene function, constructing animal models of human diseases, and genetic breeding. However, due to the low efficiency of homologous recombination, traditional homologous recombination must rely on ES cells, so it is generally limited to the application at the cellular level and in mice. In recent years, with the development and application of technologies such as ZFN (zinc finger nuclease) and TALEN (transcription activator-like effector nuclease), people can make targeted mutations to target genes in many species, but the mutations of the genes obtained by the above methods The form is random, so precise control is not possible, and knock-in and conditional knock-out are not possible.
Professor Zhang Bo's laboratory used TALEN to cause DNA double-strand breaks, which greatly improved the efficiency of homologous recombination, thereby achieving true homologous recombination gene targeting in the model organism zebrafish. All three reviewers believe that this work not only has a huge impact on the zebrafish research field, but this method should be applicable to other species, so that homologous recombination gene targeting can get rid of the dependence on ES cells, which may have a profound impact on animal husbandry Impact ("The technology may have potential to transform the practice of livestock breeding in general.").
This work is another new breakthrough after Zhang Bo Laboratories first reported internationally the use of TALEN to successfully achieve heritable genetic site-directed mutations in zebrafish in 2011. Zu Yao and Associate Professor Tong Xiangjun, the 09-level doctoral students in Professor Zhang Bo's laboratory, are co-first authors of the article. This work was supported by the 973 project and the National Natural Science Foundation of China.
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