摘要
人工miRNA(amiRNA)是一种高效的基因沉默技术,其原理是以植物内源miRNA的前体为基本骨架,将其茎环结构中的miRNA/miRNA*序列替换为amiRNA/amiRNA*序列,使产生的amiRNA作用于目标靶基因,以实现对靶基因表达的抑制。以拟南芥中转录因子基因MYB28为例,介绍了amiRNA的设计基本原理和构建方法。在MYB28基因序列的3'端、5'端和中间部分分别设计了amiRNA,以拟南芥内源的miRNA 319a前体为骨架,以靶向于MYB28的amiRNA序列替代原有的miRNA 319a序列,在拟南芥中过量表达,获得的转基因植株中MYB28表达水平平均降低了86%。根据试验结果对人工miRNA沉默基因技术的特点及应用前景进行了讨论。
Artificial miRNA（amiRNA）is an effective gene silencing technology. The basic principle of amiRNA is based on its biogenesis mechanism. Using endogenous miRNA precursor as a basic framework, miRNA/miRNA＊is then replaced by amiRNA/amiRNA＊. The produced amiRNA is expected to silence the target gene. In this paper, the basic principle and method of amiRNA construction was introduced. AmiRNAs were designed to the target gene MYB28 as an example. Three amiRNAs respectively target to the 3＇ terminal region, 5＇ region and middle region of MYB28 coding sequence were designed. Precursor of miRNA 319a in Arabidopsis thaliana was taken as a framework and miRNA319a sequence was replaced by amiRNAs targeting to MYB28 gene. The expression level of MYB28 in transgenic plants decreased 86%. Based on our experiments, the unique characteristics of amiRNA and its further applications were discussed.
出处
《生物技术通报》
CAS
CSCD
北大核心
2014年第4期50-56,共7页
Biotechnology Bulletin
基金
国家自然科学基金项目(31070265)
