Imaging of pre-mRNA splicing in living subjects using a genetically encoded luciferase reporter
Imaging of pre-mRNA splicing in living subjects using a genetically encoded luciferase reporter
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使用基因编码的荧光素酶报告基因对活体受试者中的前 mRNA 剪接进行成像
DOI：
10.1364/boe.9.000518
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发表时间：
2018-02-01
影响因子：
3.4
通讯作者：
Wang, Fu
中科院分区：
文献类型：
--
作者：
Shi, Yaru;Liu, Wei;Wang, Fu
前信使核糖核酸剪接是大多数真核生物基因表达中必不可少的一步。在这里，我们提出了一个基因编码的荧光素酶报告程序的可行性，以监测活细胞和动物中的前mRNA剪接过程。我们发现，异银杏皂苷诱导的剪接活性变化在体外可以很容易地以剂量和时间依赖的方式显示出来。此外，通过生物发光成像也可以明显检测到小鼠体内的Pre-mRNA剪接过程，并通过RT-PCR进行证实。我们的工作提供了一个报告系统，允许高通量筛选化学文库，以识别导致剪接异常模式的潜在化合物。(C)2018年OSA开放获取出版协议条款下的美国光学学会
Pre-mRNA splicing is an essential step in gene expression in most eukaryote genes. Here we present the feasibility of a genetically encoded luciferase reporter to monitor the pre-mRNA splicing process in living cells and animals. We showed that the splicing activity change induced by isoginkgetin could be readily visualized in vitro both in a dose and time dependent manner. Moreover, the pre-mRNA splicing process could be also obviously detected in mice by bioluminescence imaging and confirmed by RT-PCR. Our work provided a reporter system that allows high-throughput screening of chemical libraries to identify potential compounds leading to aberrant patterns of splicing. (C) 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement