Repositorium
High-efficiency CRISPR/Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize
171
Journal Article / 2016
Qi, Weiwei; Zhu, Tong; Tian, Zhongrui; Li, Chaobin; Zhang, Wei; Song, Rentao
Background: CRISPR/Cas9 genome editing strategy has been applied to a variety of species and the tRNA-processing system has been used to compact multiple gRNAs into one synthetic gene for manipulating multiple genes in rice. Results: We optimized and introduced the multiplex gene editing strategy based on the tRNA-processing system into maize. Maize glycine-tRNA was selected to design multiple tRNA-gRNA units for the simultaneous production of numerous gRNAs under the control of one maize U6 promoter. We designed three gRNAs for simplex editing and three multiple tRNA-gRNA units for multiplex editing. The results indicate that this system not only increased the number of targeted sites but also enhanced mutagenesis efficiency in maize. Additionally, we propose an advanced sequence selection of gRNA spacers for relatively more efficient and accurate chromosomal fragment deletion, which is important for complete abolishment of gene function especially long non-coding RNAs (lncRNAs). Our results also indicated that up to four tRNA-gRNA units in one expression cassette design can still work in maize. Conclusions: The examples reported here demonstrate the utility of the tRNA-processing system-based strategy as an efficient multiplex genome editing tool to enhance maize genetic research and breeding.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
365 |
Song, Renato China |
Zea mays |
CRISPR/Cas9 AP2 |
No information SDN1 |
Basic research Basic research |
366 |
Song, Renato China |
Zea mays |
CRISPR/Cas9 IncRNAs |
No information SDN1 |
Basic research Basic research |
367 |
Song, Renato China |
Zea mays |
CRISPR/Cas9 MADS |
Interacts with Opaque2 and activate zein gene promoters SDN1 |
Basic research Basic research |
368 |
Song, Renato China |
Zea mays |
CRISPR/Cas9 MYBR |
No information SDN1 |
Basic research Basic research |
369 |
Song, Renato China |
Zea mays |
CRISPR/Cas9 PPR |
No information SDN1 |
Basic research Basic research |
370 |
Song, Renato China |
Zea mays |
CRISPR/Cas9 RPL |
No information SDN1 |
Basic research Basic research |