Arabidopsis RNA immunoprecipitation.
Arabidopsis RNA immunoprecipitation.
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DOI：
10.1111/j.1365-313x.2009.03859.x
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发表时间：
2009-07
期刊：
影响因子：
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通讯作者：
L. Terzi;G. Simpson
中科院分区：
文献类型：
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作者：
L. Terzi;G. Simpson
RNA结合蛋白是植物基因表达的关键调控因子。与此一致的是，拟南芥基因组编码了许多RNA结合蛋白，这些蛋白在遗传上是正常发育和响应环境变化所必需的。然而，这些RNA结合蛋白控制的直接RNA靶标和RNA加工事件却知之甚少。为了便于RNA结合蛋白的功能鉴定，我们将RNA免疫沉淀实验应用于拟南芥。以U2B“-U2 SnRNA相互作用为模型实验系统，我们证明了用甲醛处理完整植物时，可以使用识别U2B”“与通用GFP标签融合的抗体来免疫捕获U2 SnRNA。与全基因组拼接阵列和高通量下一代测序的最新发展相结合，这种程序和技术的组合有可能对植物RNA结合蛋白进行系统的功能分析。
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.