Journal
PLANT JOURNAL
Volume 59, Issue 1, Pages 163-168Publisher
WILEY
DOI: 10.1111/j.1365-313X.2009.03859.x
Keywords
Arabidopsis; post-transcriptional; RNA-binding protein; RNA immunoprecipitation; RNA
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Funding
- Biotechnology and Biological Sciences Research Council [BB/D000653/1]
- Biotechnology and Biological Sciences Research Council [BB/D000653/1] Funding Source: researchfish
- BBSRC [BB/D000653/1] Funding Source: UKRI
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P>RNA-binding proteins are key regulators of plant gene expression. Consistent with this, the Arabidopsis genome encodes many RNA-binding proteins that are genetically required for normal development and for responding to environmental changes. However, the direct RNA targets and RNA processing events that these RNA-binding proteins control are poorly understood. In order to facilitate the functional characterization of RNA-binding proteins, we have applied the RNA immunoprecipitation assay to Arabidopsis. Working with the U2B '-U2 snRNA interaction as a model experimental system, we show that treatment of intact plants with formaldehyde allows immunocapture of U2 snRNA using antibodies that recognize U2B ' fused to the generic GFP tag. When coupled with recent developments in whole-genome tiling arrays and high-throughput next-generation sequencing, this combination of procedures and technology has the potential to allow systematic functional analysis of plant RNA-binding proteins.
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