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Screening and Identification of Virus-Encoded RNA Silencing Suppressors
RNA silencing, including RNA interference, is a novel method of gene regulation and one of the potent host-defense mechanisms against the viruses. In the course of evolution, the viruses have encoded proteins with the potential to suppress the host RNA silencing mechanism as a counterdefense strategy. The virus-encoded RNA silencing suppressors (RSSs) can serve as important biological tools to dissect the detailed RNA silencing pathways and also to evolve the antiviral strategies. Screening and identification of the RSSs are indeed of utmost significance in the field of plant biotechnology. We describe two Green Fluorescent Protein (GFP) reporter-based plant assay systems that rely on two different principles, namely reversal of silencing and enhancement of rolling circle replication (RCR) of geminiviral replicon. These proof-of-concept examples and assay systems could be used to screen various plant, animal, and insect viral ORFs for identification of the RSS activities.
- Cre Transgenic Mouse Lines
- Silencing of Gene Expression by Gymnotic Delivery of Antisense Oligonucleotides
- In Vitro Transcription and Translation in a Cell-Free System from Clostridium tetani
- Purification of Restriction Fragments Containing Replication Intermediates from Complex Genomes for 2-D Gel Analysis
- Molecular Analysis of Chromosome Aberrations in Hematological Malignancies: Southern Hybridization
- Two-Stage Polymerase Chain Reaction Protocol Allowing Introduction of Multiple Mutations, Deletions, and Insertions, Using QuikC
- A PCR-Based Method for Studying DNA Methylation
- Site-Directed Mutagenesis Mediated by a Single Polymerase Chain Reaction Product
- Using the Quantitative Competitive RT-PCR Technique to Analyze Minute Amounts of Different mRNAs in Small Tissue Samples
- Identification of Viral Suppressors of RNAi by a Reporter Assay in Drosophila S2 Cell Culture
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