Challenges to increasing targeting efficiency in genome engineering |
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Authors: | Takuro HORII Izuho HATADA |
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Institution: | 1)Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Gunma 371-8512, Japan |
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Abstract: | Gene targeting technologies are essential for the analysis of gene functions. Knockout mouse generation via
genetic modification of embryonic stem cells (ESCs) is the commonest example, but it is a time-consuming and
labor-intensive procedure. Recently, a novel genome editing technology called CRISPR/Cas has enabled the
direct production of knockout mice by non-homologous end joining (NHEJ)-mediated mutations. Unexpectedly,
however, it generally exhibits a low efficiency in homologous recombination (HR) and is prone to high
mosaicism. Meanwhile, gene targeting using ESCs is still being improved, as reported by Fukuda et
al. in this issue. Here, we outline current gene targeting technologies with special emphasis on
HR-mediated technologies, which are currently being performed using these two major strategies. |
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Keywords: | Gene targeting Genome editing Homologous recombination Knockin Knockout |
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