Wesley Susan Varsha, Liu Qing, Wielopolska Anna, Ellacott Geoff, Smith Neil, Singh Surinder, Helliwell Chris
CSIRO Plant Industry, Canberra, Australia.
Methods Mol Biol. 2003;236:273-86. doi: 10.1385/1-59259-413-1:273.
Hairpin (hpRNA)-mediated gene silencing exploits a cellular mechanism that recognizes double-stranded RNA (dsRNA) and subjects it and its corresponding mRNA to a sequence-specific degradation. This phenomenon is known as posttranscriptional gene silencing (PTGS) in plants and RNA interference (RNAi) in animals. dsRNA, when introduced into plant cells through hpRNA constructs, results in severe reduction of the target mRNA-the silencing effect being stably inherited over many generations. While hpRNA constructs can be made using conventional plasmids, use of generic vectors such as pHANNIBAL makes it more convenient to silence a number of genes simultaneously. Vectors, such as pHELLSGATE, that are based on the Gateway(R) technology are suitable for high-throughput gene silencing. The specificity of dsRNA silencing, it's ability to simultaneously silence multiple genes combined with the availability of high-throughput silencing vectors enables the researcher to generate custom knockout plants.
发夹（hpRNA）介导的基因沉默利用了一种细胞机制，该机制识别双链RNA（dsRNA）并使其及其相应的mRNA发生序列特异性降解。这种现象在植物中被称为转录后基因沉默（PTGS），在动物中被称为RNA干扰（RNAi）。当通过hpRNA构建体将dsRNA导入植物细胞时，会导致目标mRNA严重减少——这种沉默效应可在许多代中稳定遗传。虽然可以使用常规质粒构建hpRNA构建体，但使用诸如pHANNIBAL等通用载体可以更方便地同时沉默多个基因。基于Gateway®技术的载体，如pHELLSGATE，适用于高通量基因沉默。dsRNA沉默的特异性、同时沉默多个基因的能力以及高通量沉默载体的可用性，使研究人员能够生成定制的基因敲除植物。
