Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas-Universidad Politécnica de Valencia), Valencia, Spain.
Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy.
Methods Mol Biol. 2022;2408:227-242. doi: 10.1007/978-1-0716-1875-2_15.
RNAi-based tools are widely used in gene function studies and for crop improvement. However, no effective methods for precisely controlling the degree of induced silencing have been reported until recently. Here we report a detailed protocol for designing and generating synthetic trans-acting small interfering RNA (syn-tasiRNA) constructs for fine-tuning gene expression in plants. Recently developed high-throughput AtTAS1c-D2-B/c-based vectors are used to clone and express syn-tasiRNAs that possess different efficacies depending on their precursor location and on their degree of base-pairing with the 5' end of target RNAs.
基于 RNAi 的工具广泛用于基因功能研究和作物改良。然而，直到最近才报道了一种有效方法来精确控制诱导沉默的程度。在这里，我们报告了一种用于设计和生成用于微调植物中基因表达的合成反式作用小干扰 RNA（syn-tasiRNA）构建体的详细方案。最近开发的基于高通量 AtTAS1c-D2-B/c 的载体用于克隆和表达 syn-tasiRNAs，这些 syn-tasiRNAs 的功效取决于它们的前体位置以及与靶 RNA5' 端的碱基配对程度。
Zotero 插件