Using Nuclear Run-On Transcription Assays in RNAi Studies
RNA interference (RNAi) is a mechanism regulating gene transcript levels either by transcriptional gene silencing or by posttranscriptional gene silencing, which act in the genome maintenance and the regulation of gene expression which is typically inferred from measuring transcript abundance. Nuclear “run-on” (or “run-off”) transcription assays have been used to obtain quantitative information about the relative rates of transcription of different genes in nuclei isolated from a particular tissue or organ. Basically, these assays exploit the activity of RNA polymerases to synthesize radiolabeled transcripts that then can be hybridized to filter-bound, cold, excess single-stranded DNA probes representing genes of interest. The protocol presented here streamlines, adapts, and optimizes nuclear run-on transcription assays for use in RNAi studies.
- Immunological Screening of -Phage cDNA Expression Libraries
- Advances in RIP-Chip Analysis: RNA-Binding Protein Immunoprecipitation-Microarray Profiling
- Modeling and Analysis of ChIP-Chip Experiments
- HIV-Based Vectors: Preparation and Use
- Herpes Simplex Virus-Cell Interactions Studied by Immunogold Cryosection Electron Microscopy
- Application of the Firefly Luciferase Reporter Gene
- Hematopoietic Stem and Progenitor Cells
- Site-Directed Mutagenesis of Double-Stranded Plasmids, Domain Substitution, and Marker Rescue by Comutagenesis of Restriction En
- Ischemic Stroke in Mice and Rats
- RNA Refolding Studied by Light-Coupled NMR Spectroscopy