Abstract
Genome editing technologies such as ZFN, TALEN, and CRISPR/Cas9 efficiently induce DNA double-stranded breaks (DSBs) at a targeted genomic locus, often resulting in a frameshift-mediated target gene disruption. It remains difficult to perform targeted integration of exogenous genes by genome editing technologies. DSBs can be restored through DNA repair mechanisms, such as non-homologous end joining (NHEJ), microhomology-mediated end joining (MMEJ), and homologous recombination (HR). It is well known that HR facilitates homology-dependent integration of donor DNA template into a targeted locus. Recently, both NHEJ-mediated and MMEJ-mediated targeted integrations of exogenous genes have been developed in zebrafish. This chapter summarizes the application of CRISPR/Cas9-mediated knock-in technology in zebrafish.
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Acknowledgment
This work was supported by the Japan Society for the Promotion of Science (JSPS, 26640064), Japan Agency for Medical Research and Development (AMED, 16km0210077j0001), the Takeda Science Foundation, and the SENSHIN Medical Research Foundation.
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Kawahara, A. (2017). CRISPR/Cas9-Mediated Targeted Knockin of Exogenous Reporter Genes in Zebrafish. In: Hatada, I. (eds) Genome Editing in Animals. Methods in Molecular Biology, vol 1630. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7128-2_14
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DOI: https://doi.org/10.1007/978-1-4939-7128-2_14
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