Wong Gigi, Millar Anthony A
Research School of Biology, Plant Science Division, Australian National University, Canberra, ACT, Australia.
Methods Mol Biol. 2019;1932:227-238. doi: 10.1007/978-1-4939-9042-9_17.
The study of gene function is best achieved through the generation of loss-of-function mutants. However, for many plant microRNAs (miRNAs), this has proven challenging, as they often belong to sequence-related families, which are encoded by multiple genes that are functionally redundant. To overcome this issue, transgenic methods have been developed that express miRNA decoys, which can sequester and inhibit families of sequence-related miRNAs. This includes miRNA MIMICs, SHORT TANDEM TARGET MIMICs, and miRNA SPONGEs. Here, we describe the methods to generate transgenic Arabidopsis that express these miRNA decoys in order to determine miRNA function.
通过产生功能缺失突变体来研究基因功能是最佳途径。然而，对于许多植物微小RNA（miRNA）而言，这已被证明具有挑战性，因为它们通常属于序列相关家族，由多个功能冗余的基因编码。为克服这一问题，已开发出表达miRNA诱饵的转基因方法，这些诱饵可以隔离并抑制序列相关的miRNA家族。这包括miRNA模拟物、短串联靶标模拟物和miRNA海绵。在此，我们描述了生成表达这些miRNA诱饵的转基因拟南芥以确定miRNA功能的方法。
