Homology-based double-strand break-induced genome engineering in plants
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This review summarises the recent progress in DSB-induced gene targeting by homologous recombination in plants. We are getting closer to efficiently inserting genes or precisely exchanging single amino acids.
Although the basic features of double-strand break (DSB)-induced genome engineering were established more than 20 years ago, only in recent years has the technique come into the focus of plant biologists. Today, most scientists apply the recently discovered CRISPR/Cas system for inducing site-specific DSBs in genes of interest to obtain mutations by non-homologous end joining (NHEJ), which is the prevailing and often imprecise mechanism of DSB repair in somatic plant cells. However, predefined changes like the site-specific insertion of foreign genes or an exchange of single amino acids can be achieved by DSB-induced homologous recombination (HR). Although DSB induction drastically enhances the efficiency of HR, the efficiency is still about two orders of magnitude lower than that of NHEJ. Therefore, significant effort have been put forth to improve DSB-induced HR based technologies. This review summarises the previous studies as well as discusses the most recent developments in using the CRISPR/Cas system to improve these processes for plants.
KeywordsDouble-strand break repair Homologous recombination Non-homologous end joining Synthetic nucleases Targeted mutagenesis Gene targeting
We apologise to all colleagues in this field for not being able to cite all of the rapidly growing number of reports published on genome engineering in plants due to space limitations. Work on DSB-induced genome engineering in our lab has been funded by the Deutsche Forschungsgemeinschaft, the Bundesministerium für Bildung und Forschung (BMBF), the EU and the ERC (Advanced Grant COMREC).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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