Base editing in tomatoes was achieved by transient expression.
The Solanaceae plants, particularly the tomato (Solanum lycopersicum), is of huge economic value worldwide. The tomato is a unique model plant for studying the functions of genes related to fruit ripening. Deeper understanding of tomatoes is of great importance for both plant research and the economy. Genome editing technology, such as CRISPR/Cas9, has been used for functional genetic research. However, some challenges, such as low transformation efficiency, remain with this technology. Moreover, the foreign Cas9 and gRNA expression cassettes must be removed to obtain null-segregants In this study, we used a high-level transient expression system to improve the base editing technology. A high-level transient expression system has been established previously using geminiviral replication and a double terminator. The pBYR2HS vector was used for this transient expression system. nCas9-CDA and sgRNA-SlHWS were introduced into this vector, and the protein and RNA were then transiently expressed in tomato tissues by agroinfiltration. The homozygous mutant produced by base editing was obtained in the next generation with an efficiency of about 18%. nCas9-free next-generation plants were 71%. All the homozygous base-edited plants in next generation are nCas9-free. These findings show that the high-level transient expression system is useful for base editing in tomatoes.
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Activation-induced cytidine deaminase
Bean yellow dwarf virus
CRISPR-associated endonuclease 9
Clustered regularly interspaced short palindromic repeats
Double strand break
Insertion and/or deletion
Long intergenic region
Single guide RNA
Solanum lycopersicum HAWAIIAN SKIRT
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We thank Ms. Kazuko Ito and Ms. Yuri Nemoto at Tsukuba-Plant Innovation Research Center (T-PIRC), University of Tsukuba for technical support. Tomato seeds (cv. Micro-Tom, TOMJPF00001) were provided from T-PIRC, University of Tsukuba through the National Bio-Resource Project (NBRP) of the Japan Agency for Research and Development (AMED), Japan. This work was supported by the Cross-ministerial Strategic Innovation Promotion Program (SIP), Japan, Program on Open Innovation Platform with Enterprises, Research Institute and Academia, Japan Science and Technology Agency (JST, OPERA, JPMJOP1851), and a Cooperative Research Grant from the Plant Transgenic Design Initiative, Gene Research Center, T-PIRC, University of Tsukuba.
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Communicated by Hiroyasu Ebinuma.
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Yuan, S., Kawasaki, S., Abdellatif, I.M.Y. et al. Efficient base editing in tomato using a highly expressed transient system. Plant Cell Rep (2021). https://doi.org/10.1007/s00299-021-02662-z
- Transient expression
- Tsukuba system
- Solanum lycopersicum