The DOI matches its Crossref record; 28 of the 28 references it cites were found in a registry or an index.
Comparison of CRISPR/Cas9 expression constructs for efficient targeted mutagenesis in rice
Open accessAbstract
The CRISPR/Cas9 system is an efficient tool used for genome editing in a variety of organisms. Despite several recent reports of successful targeted mutagenesis using the CRISPR/Cas9 system in plants, in each case the target gene of interest, the Cas9 expression system and guide-RNA (gRNA) used, and the tissues used for transformation and subsequent mutagenesis differed, hence the reported frequencies of targeted mutagenesis cannot be compared directly. Here, we evaluated mutation frequency in rice using different Cas9 and/or gRNA expression cassettes under standardized experimental conditions. We introduced Cas9 and gRNA expression cassettes separately or sequentially into rice calli, and assessed the frequency of mutagenesis at the same endogenous targeted sequences. Mutation frequencies differed significantly depending on the Cas9 expression cassette used. In addition, a gRNA driven by the OsU6 promoter was superior to one driven by the OsU3 promoter. Using an all-in-one expression vector harboring the best combined Cas9/gRNA expression cassette resulted in a much improved frequency of targeted mutagenesis in rice calli, and bi-allelic mutant plants were produced in the T0 generation. The approach presented here could be adapted to optimize the construction of Cas9/gRNA cassettes for genome editing in a variety of plants.
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Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
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Multiplex Genome Engineering Using CRISPR/Cas Systems
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RNA-Guided Human Genome Engineering via Cas9
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Efficient genome editing in zebrafish using a CRISPR-Cas system
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Cited by
Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion
Shimatani, Z., Kashojiya, S., Takayama, M., Terada, R., Arazoe, T., Ishii, H., Teramura, H., Yamamoto, T., Komatsu, H., Miura, K., et al. (2017) · Nature Biotechnology · doi:10.1038/nbt.3833 · Cited by 812
CRISPR/Cas9‐mediated efficient targeted mutagenesis in grape in the first generation
Wang, X., Tu, M., Wang, D., Liu, J., Li, Y., Li, Z., Wang, Y., & Wang, X. (2018) · Plant Biotechnology Journal · doi:10.1111/pbi.12832 · Cited by 367
Efficient increase of ɣ-aminobutyric acid (GABA) content in tomato fruits by targeted mutagenesis
Nonaka, S., Arai, C., Takayama, M., Matsukura, C., & Ezura, H. (2017) · Scientific Reports · doi:10.1038/s41598-017-06400-y · Cited by 357
Shen, C., Que, Z., Xia, Y., Tang, N., Li, D., He, R., & Cao, M. (2017) · Journal of Plant Biology · doi:10.1007/s12374-016-0400-1 · Cited by 234
Genome Editing Weds CRISPR: What Is in It for Phytoremediation?
Basharat, Z., Novo, L. A. B., & Yasmin, A. (2018) · Plants · doi:10.3390/plants7030051 · Cited by 104
Metadata
- Authors
- Mikami, M., Toki, S., & Endo, M.
- Venue
- Plant Molecular Biology
- Published
- 2015-07-19
- Volume
- 88
- Issue
- 6
- Pages
- 561-572
- ISSN
- 0167-4412 (ISSN-L)
- 1573-5028
- OpenAlex
- W1885181197
- Cited by
- 296
- References
- 28
- FWCI
- 9.13
- Type
- journal article
- Language
- en
- Open access
- hybrid
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