Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Generation of Transgenic Arabidopsis
4.2. dsRNA Synthesis and Application
4.3. Total RNA Extraction, Reverse Transcription, and qRT-PCR
4.4. Laser Scanning Microscopy
4.5. Protein Extraction and Western Blotting
4.6. DNA Extraction and Bisulfite Sequencing
4.7. Detection of EGFP-siRNA by Stem-Loop RT-PCR and DNA Sequencing
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene | Treatment | CG | CHG | CHH |
---|---|---|---|---|
NPTII | Water (before) Water (7 dpa) | 65.6 ± 2.1 | 60.1 ± 3.3 | 55.9 ± 3.8 |
65.3 ± 3.2 | 63.3 ± 2.4 | 59.3 ± 4.7 | ||
NPTII-dsRNA (before) NPTII-dsRNA (7 dpa) | 68.5 ± 4.4 | 60.7 ± 3.0 | 51.6 ± 4.5 | |
82.5 ± 4.9 ** | 79.2 ± 3.4 ** | 77.5 ± 3.6 ** | ||
EGFP-dsRNA (before) EGFP-dsRNA (7 dpa) | 65.3 ± 3.4 | 65.0 ± 3.7 | 55.6 ± 2.4 | |
80.7 ± 4.3 | 78.2 ± 2.9 | 78.1 ± 2.2 ** | ||
EGFP | Water (before) Water (7 dpa) | 62.1 ± 2.4 | 59.2 ± 3.1 | 54.7 ± 3.9 |
62.4 ± 4.2 | 59.5 ± 2.8 | 57.9 ± 5.2 | ||
NPTII-dsRNA (before) NPTII-dsRNA (7 dpa) | 65.2 ± 5.9 | 62.2 ± 3.3 | 59.3 ± 4.3 | |
75.8 ± 5.7 | 75.9 ± 4.6 * | 73.9 ± 3.4 ** | ||
EGFP-dsRNA (before) EGFP-dsRNA (7 dpa) | 59.9 ± 4.2 | 56.1 ± 3.8 | 54.1 ± 3.2 | |
74.9 ± 4.8 | 75.1 ± 4.7 ** | 73.1 ± 3.0 ** |
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Dubrovina, A.S.; Aleynova, O.A.; Kalachev, A.V.; Suprun, A.R.; Ogneva, Z.V.; Kiselev, K.V. Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA. Int. J. Mol. Sci. 2019, 20, 1585. https://doi.org/10.3390/ijms20071585
Dubrovina AS, Aleynova OA, Kalachev AV, Suprun AR, Ogneva ZV, Kiselev KV. Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA. International Journal of Molecular Sciences. 2019; 20(7):1585. https://doi.org/10.3390/ijms20071585
Chicago/Turabian StyleDubrovina, Alexandra S., Olga A. Aleynova, Alexander V. Kalachev, Andrey R. Suprun, Zlata V. Ogneva, and Konstantin V. Kiselev. 2019. "Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA" International Journal of Molecular Sciences 20, no. 7: 1585. https://doi.org/10.3390/ijms20071585