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Screening and Identification of RNA Silencing Suppressors from Secreted Effectors of Plant Pathogens
Published on: February 3, 2020
Local RNA silencing mediated by agroinfiltration
Jutta Maria Helm1, Elena Dadami, Kriton Kalantidis
1Institute of Molecular Biology and Biotechnology, Heraklion, Greece.
Methods in Molecular Biology (Clifton, N.J.)
|May 3, 2011
Summary
Agroinfiltration is a rapid plant gene expression technique. This method effectively induces local and systemic gene silencing, with protocols for assay evaluation provided.
Area of Science:
- Plant Molecular Biology
- Gene Expression Analysis
- RNA Interference (RNAi)
Background:
- Agroinfiltration offers rapid, transient in planta gene expression.
- It is valuable for short-term protein overexpression studies.
- Agroinfiltration has shown significant success in inducing RNA silencing.
Purpose of the Study:
- To detail the application of agroinfiltration for inducing gene silencing in plants.
- To describe protocols for local and systemic transgene silencing.
- To outline methods for evaluating RNA silencing assay efficiency.
Main Methods:
- Utilizing agroinfiltration for transient gene expression in plants.
- Inducing local silencing of endogenous genes and transgenes.
- Achieving systemic silencing of transgenes.
- Performing co-infiltration assays for gene expression studies.
- Detecting small interfering RNAs (siRNAs) to confirm RNA silencing.
- Quantifying target sequence effects to measure assay efficiency.
Main Results:
- Demonstrated successful induction of local gene silencing via agroinfiltration.
- Showcased the capability of agroinfiltration to induce systemic gene silencing.
- Established protocols for evaluating the efficiency of RNA silencing assays.
- Validated the detection of characteristic siRNAs and target sequence modulation.
Conclusions:
- Agroinfiltration is a versatile and efficient tool for transient gene expression and RNA silencing in plants.
- The described protocols enable robust induction and evaluation of both local and systemic gene silencing.
- This method facilitates further research in gene function and regulation through RNA interference.
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