CRISPR/Cas9-Mediated Deletion of Large Genomic Fragments in Soybean
Abstract
:1. Introduction
2. Results
2.1. Experimental Design of Targeted Fragment Deletions of GmFT2a and GmFT5a
2.2. Detection of Site-Specific Mutations and Large Fragment Deletions
2.3. Inheritance of Large Fragment Deletion Mutations and Phenotypes of the Mutants
3. Discussion
4. Materials and Methods
4.1. SgRNA Design and Construction of the Combinatory Dual-sgRNA/Cas9 Vectors
4.2. Determination of the Site-Specific Mutation Frequencies at Each Target Site and Large Fragment Deletions
4.3. Flowering Time Measurements and Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Target Sites | No. of Transgenic Events Identified | No. of Events with Targeted Mutations | Mutation Frequency |
---|---|---|---|
GmFT2a-D1 | 25 | 12 | 48.0% |
GmFT2a-D2 | 21 | 11 | 52.4% |
GmFT2a-D3 | 18 | 13 | 72.2% |
GmFT5a-D1 | 24 | 17 | 70.8% |
GmFT5a-D2 | 14 | 5 | 35.7% |
Constructs | No. of Transgenic Plants Identified a | No. of Plants with no Mutation | No. of Plants with Targeted Mutations at the First Site | No. of Plants with Targeted Mutations at the Second Site | No. of Plants with Simultaneous Mutations at the Two Target Sites | No. of Plants with Large Fragment Deletions |
---|---|---|---|---|---|---|
GmFT2a-D1/D2 | 32 | 11 (34.4%) | 16 (50.0%) | 15 (46.9%) | 10 (31.2%) | 5 (15.6%) |
GmFT2a-D1/D3 | 33 | 11 (33.3%) | 17 (51.5%) | 19 (57.6%) | 14 (42.4%) | 4 (12.1%) |
GmFT5a-D1/D2 | 19 | 7 (36.8%) | 12 (63.2%) | 7 (36.8%) | 7 (36.8%) | 3 (15.8%) |
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Cai, Y.; Chen, L.; Sun, S.; Wu, C.; Yao, W.; Jiang, B.; Han, T.; Hou, W. CRISPR/Cas9-Mediated Deletion of Large Genomic Fragments in Soybean. Int. J. Mol. Sci. 2018, 19, 3835. https://doi.org/10.3390/ijms19123835
Cai Y, Chen L, Sun S, Wu C, Yao W, Jiang B, Han T, Hou W. CRISPR/Cas9-Mediated Deletion of Large Genomic Fragments in Soybean. International Journal of Molecular Sciences. 2018; 19(12):3835. https://doi.org/10.3390/ijms19123835
Chicago/Turabian StyleCai, Yupeng, Li Chen, Shi Sun, Cunxiang Wu, Weiwei Yao, Bingjun Jiang, Tianfu Han, and Wensheng Hou. 2018. "CRISPR/Cas9-Mediated Deletion of Large Genomic Fragments in Soybean" International Journal of Molecular Sciences 19, no. 12: 3835. https://doi.org/10.3390/ijms19123835