Plant Gene Silencing Service
Loss-of-function (LOF) experiments are a cornerstone of our understanding of plant biological processes. While gene knockout has long been considered the "gold standard" for LOF research, it does not apply to the high-copy genes in plants. Therefore, CD BioSciences offers RNA interference (RNAi) services, including post-transcriptional gene silencing (PTGS) and transcriptional gene silencing (TGS) technologies to suppress endogenous gene expression in plants.
What is RNAi?
RNAi technology introduces double-stranded RNA (dsRNA) into plants. The long-stranded dsRNA is cleaved by a Dicer into small interfering RNAs or siRNAs of 21-25 nucleotides. siRNAs then assemble proteins into RNA-induced silencing complexes (RISCs). The activated RISC binds to the complementary transcript, causing the bound mRNA to be cleaved and degraded, resulting in gene silencing.
Figure 1. Schematic diagram of the main RNAi pathways used by plants to silence target genes. (Ashfaq, M. A., et al. 2020) Service Content
Design siRNA target sequence
- Selection of siRNA target sites.
- Sequence homology analysis.
- Design negative control.
Labelling of siRNA
To better detect the transfection efficiency of siRNA and locate the siRNA, we can label the synthetic siRNA, our labelling methods include
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Selection of RNAi expression vectors
By cloning the dsDNA template sequence corresponding to the siRNA into the back of the promoter of RNA polymerase III, we can express the desired siRNA molecule in vivo. It is possible to achieve a longer gene silencing effect without direct manipulation of RNA.
Synthesis and validation of siRNA
We provide siRNA synthesis methods including
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After synthesising the siRNA, the target gene interference efficiency is detected using qRT-PCR or Western Blotting to verify whether the synthesised siRNA is effective.
RNAi induction service in plants
| Our technologies | Details |
| Micro bombardment | Microinjection of dsRNA or intron-containing hairpin structure RNA (iphRNA) expression vectors into plants. |
| Agrobacterium transformation | T-DNA plasmids with exogenous genes that can be transcribed into ihpRNAs are integrated into plant genomes through Agrobacterium-mediated integration. |
| Virus-induced gene silencing (VIGS) | VIGS inhibits the expression of endogenous genes in plants by inserting recombinant viruses into target gene fragments. It can silence target genes in infected plants and avoid plant transformation. |
Application Fields
- RNAi-induced gene silencing is an effective tool for designing disease-resistant plant lines against diseases caused by bacteria, fungi, and viruses.
- RNAi can effectively defend against insects and parasites, opening up new avenues for crop protection.
- As a new approach, RNAi has greater potential for crop quality traits and nutritional improvement by altering the expression of plant genes.
- RNAi plays an essential role in abiotic stress resistance in crops.
CD BioSciences is dedicated to advancing plant science and biotechnology by unlocking the potential of gene-silencing mechanisms. Through precise and efficient gene silencing technologies, we can inhibit specific gene expression, enabling researchers to study gene function, elucidate complex biological pathways, and develop innovative solutions for crop improvement. If you have requirements for exploring plant gene function, please contact us for more information.
References
- Ashfaq, M. A., et al. (2020). Post-transcriptional gene silencing: Basic concepts and applications. Journal of Biosciences,.45(1),128.
- Guo, Q., et al. (2016). RNA Silencing in Plants: Mechanisms, Technologies and Applications in Horticultural Crops. Current genomics. 17(6), 476-489.
- El-Sappah, A. H., et al. (2021). Comprehensive Mechanism of Gene Silencing and Its Role in Plant Growth and Development. Frontiers in plant science. 12, 705249.
For research use only, not for clinical use.