Precision genome editing in plants via gene targeting and piggy B ac ‐mediated marker excision
| Autor: | Namie Ohtsuki, Hiroaki Saika, Ayako Nishizawa-Yokoi, Seiichi Toki, Masaki Endo |
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| Rok vydání: | 2014 |
| Předmět: |
Transposable element
Transposases Oryza sativa Plant Science Biology Genome gene targeting Genome engineering Genome editing Genetics Point Mutation Gene Selectable marker Transposase marker excision Herbicides piggyBac transposon food and beverages Gene targeting Oryza Cell Biology Plants Genetically Modified Acetolactate Synthase cleistogamy 1 Technical Advance Mutagenesis Genetic Engineering Genome Plant Herbicide Resistance |
| Zdroj: | The Plant Journal |
| ISSN: | 1365-313X 0960-7412 |
| Popis: | Precise genome engineering via homologous recombination (HR)-mediated gene targeting (GT) has become an essential tool in molecular breeding as well as in basic plant science. As HR-mediated GT is an extremely rare event, positive-negative selection has been used extensively in flowering plants to isolate cells in which GT has occurred. In order to utilize GT as a methodology for precision mutagenesis, the positive selectable marker gene should be completely eliminated from the GT locus. Here, we introduce targeted point mutations conferring resistance to herbicide into the rice acetolactate synthase (ALS) gene via GT with subsequent marker excision by piggyBac transposition. Almost all regenerated plants expressing piggyBac transposase contained exclusively targeted point mutations without concomitant re-integration of the transposon, resulting in these progeny showing a herbicide bispyribac sodium (BS)-tolerant phenotype. This approach was also applied successfully to the editing of a microRNA targeting site in the rice cleistogamy 1 gene. Therefore, our approach provides a general strategy for the targeted modification of endogenous genes in plants. |
| Databáze: | OpenAIRE |
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