Wang Fang, Qiao Huan-Huan, Xu Rong-Gang, Sun Jin, Zhu Ruibao, Mao Decai, Ni Jian-Quan
Gene Regulatory Lab, School of Medicine, Tsinghua University, Beijing 100084, China.
Tsingdao Advanced Research Institute, Tongji University, Qingdao 266000, China.
Bio Protoc. 2019 Feb 5;9(3):e3158. doi: 10.21769/BioProtoc.3158.
Much of our knowledge about the mechanisms underlying biological processes relies on genetic approaches, whereby gene activity is reduced and the phenotypic consequences of perturbation are analyzed in detail. For functional genomic studies, a specific, systematic, and cost-effective manner is critical. Transgenic RNAi system is the top priority choice to study gene functions due to its simple and practical characteristics in . We established a novel system that works well in both soma and germ cells which is efficient and specific. With this system, we can precisely and efficiently modulate highly expressed genes, and simultaneously knock down multiple genes in one step. In this study, we provide a detailed protocol of the pNP system, which replaces other transgenic systems, and expect it can provide some help to researchers who are using this system.
我们对生物过程潜在机制的许多了解都依赖于遗传学方法,即通过降低基因活性并详细分析干扰的表型后果。对于功能基因组学研究而言,一种特定、系统且具有成本效益的方式至关重要。转基因RNAi系统因其在……方面简单实用的特性,是研究基因功能的首要选择。我们建立了一种在体细胞和生殖细胞中均能良好发挥作用的新型系统,该系统高效且特异。借助这个系统,我们能够精确且高效地调控高表达基因,并能在一步操作中同时敲低多个基因。在本研究中,我们提供了pNP系统的详细方案,该系统可替代其他转基因系统,并期望它能为正在使用该系统的研究人员提供一些帮助。