Morita Sumiyo, Horii Takuro, Kimura Mika, Kobayashi Ryosuke, Tanaka Hiroki, Akita Hidetaka, Hatada Izuho
Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan.
Laboratory of DDS Design and Drug Disposition, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Int J Mol Sci. 2023 Sep 19;24(18):14299. doi: 10.3390/ijms241814299.
Knockout mice are useful tools that can provide information about the normal function of genes, including their biochemical, developmental, and physiological roles. One problem associated with the generation of knockout mice is that the loss of some genes of interest produces a lethal phenotype. Therefore, the use of conditioned knockout mice, in which genes are disrupted in specific organs, is essential for the elucidation of disease pathogenesis and the verification of drug targets. In general, conditional knockout mice are produced using the Cre/loxP system; however, the production of the large numbers of Cre/flox knockout and control mice required for analysis requires substantial time and effort. Here, we describe the generation of liver-specific conditional knockout mice via the introduction of lipid nanoparticles encapsulating mRNA into the liver of floxed mice. This technique does not require the production of offspring by mating floxed mice and is therefore more convenient than the conventional method. The results presented here demonstrate that the LNP-based method enables liver-specific gene knockout in a short period of time.
基因敲除小鼠是有用的工具，可提供有关基因正常功能的信息，包括其生化、发育和生理作用。与基因敲除小鼠的产生相关的一个问题是，某些感兴趣的基因缺失会产生致死表型。因此，使用条件性基因敲除小鼠（其中基因在特定器官中被破坏）对于阐明疾病发病机制和验证药物靶点至关重要。一般来说，条件性基因敲除小鼠是使用Cre/loxP系统产生的；然而，分析所需的大量Cre/flox基因敲除小鼠和对照小鼠的产生需要大量的时间和精力。在这里，我们描述了通过将包裹mRNA的脂质纳米颗粒引入floxed小鼠的肝脏来产生肝脏特异性条件性基因敲除小鼠。该技术不需要通过floxed小鼠交配产生后代，因此比传统方法更方便。这里呈现的结果表明，基于脂质纳米颗粒的方法能够在短时间内实现肝脏特异性基因敲除。
