The DOI matches its Crossref record; 7 of the 7 references it cites were found in a registry or an index.
Targeted mutagenesis in soybean using the CRISPR-Cas9 system
Open accessAbstract
Genome editing is a valuable technique for gene function analysis and crop improvement. Over the past two years, the CRISPR-Cas9 system has emerged as a powerful tool for precisely targeted gene editing. In this study, we predicted 11 U6 genes in soybean (Glycine max L.). We then constructed two vectors (pCas9-GmU6-sgRNA and pCas9-AtU6-sgRNA) using the soybean U6-10 and Arabidopsis U6-26 promoters, respectively, to produce synthetic guide RNAs (sgRNAs) for targeted gene mutagenesis. Three genes, Glyma06g14180, Glyma08g02290 and Glyma12g37050, were selected as targets. Mutations of these three genes were detected in soybean protoplasts. The vectors were then transformed into soybean hairy roots by Agrobacterium rhizogenes infection, resulting in efficient target gene editing. Mutation efficiencies ranged from 3.2–9.7% using the pCas9-AtU6-sgRNA vector and 14.7–20.2% with the pCas9-GmU6-sgRNA vector. Biallelic mutations in Glyma06g14180 and Glyma08g02290 were detected in transgenic hairy roots. Off-target activities associated with Glyma06g14180 and Glyma12g37050 were also detected. Off-target activity would improve mutation efficiency for the construction of a saturated gene mutation library in soybean. Targeted mutagenesis using the CRISPR-Cas9 system should advance soybean functional genomic research, especially that of genes involved in the roots and nodules.
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Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
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DNA targeting specificity of RNA-guided Cas9 nucleases
Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann, S., Agarwala, V., Li, Y., Fine, E. J., Wu, X., Shalem, O., et al. (2013) · Nature Biotechnology · doi:10.1038/nbt.2647 · Cited by 4,741
High-Frequency off-Target Mutagenesis Induced by CRISPR-Cas Nucleases in Human Cells
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CRISPR RNA Maturation by Trans-Encoded Small RNA and Host Factor RNase III
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Cited by
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
Enhancing genetic gain in the era of molecular breeding
Xu, Y., Li, P., Zou, C., Lu, Y., Xie, C., Zhang, X., Prasanna, B. M., & Olsen, M. S. (2017) · Journal of Experimental Botany · doi:10.1093/jxb/erx135 · Cited by 336
Genome editing in cotton with the CRISPR/cas9 system
Gao, W., Long, L., Tian, X., Xu, F., Liu, J., Singh, P. K., Botella, J. R., & Song, C. (2017) · Frontiers in Plant Science · doi:10.3389/fpls.2017.01364 · Cited by 254
The CRISPR/Cas Genome-Editing Tool: Application in Improvement of Crops
Khatodia, S., Bhatotia, K., Passricha, N., Khurana, S. M. P., & Tuteja, N. (2016) · Frontiers in Plant Science · doi:10.3389/fpls.2016.00506 · Cited by 267
Yang, H., Wu, J.-J., Tang, T., Liu, K.-D., & Dai, C. (2017) · Scientific Reports · doi:10.1038/s41598-017-07871-9 · Cited by 199
Metadata
- Authors
- Sun, X., Hu, Z., Chen, R., Jiang, Q., Song, G., Zhang, H., & Xi, Y.
- Venue
- Scientific Reports
- Published
- 2015-05-29
- Volume
- 5
- Issue
- 1
- Pages
- 10342-10342
- ISSN
- 2045-2322 (ISSN-L)
- OpenAlex
- W1510606824
- Cited by
- 371
- References
- 7
- FWCI
- 11.96
- Type
- journal article
- Language
- en
- Open access
- gold
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