Creating conditional dual fluorescence labeled transgenic animals for studying function of small noncoding RNAs.
Creating conditional dual fluorescence labeled transgenic animals for studying function of small noncoding RNAs.
复制标题
创建有条件的双荧光标记转基因动物，用于研究小非编码 RNA 的功能。
DOI：
10.1080/03008207.2016.1247834
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发表时间：
2017
影响因子：
2.9
通讯作者：
Chen,Qian
中科院分区：
文献类型：
--
作者：
Yang,Kun;Gao,Yun;Yang,Mingfu;Xu,Zuoshang;Chen,Qian
Because the function of most noncoding (nc) RNAs is unknown, Cre-lox transgenic mice are useful tools to determine their functions in a tissue or developmental stage-specific manner. However, the technology faces challenges because expression of ncRNA-transgene lacks protein product. No antibody or peptide-tag can be used to trace ncRNA expression in mouse tissues in real time. Furthermore, transgene integration at different locus or orientations in the genome may result in recombination of genomic fragments in the Cre-lox system. Establishing a reliable method that can be used to determine the precise copy number and orientation of the transgene is critical to the field. We developed a fast and straightforward method to determine ncRNA-transgene copy number, orientation, and insertion site in the genome. Furthermore, upon tissue-specific expression of ncRNA, a Cre-loxP-mediated dual-fluorescence expression system facilitates fluorescence signal switching from green to red, which enables real-time monitoring of ncRNA expression by fluorescence signals. As proof of concept, we demonstrate that after microRNA (miRNA)-Flox mice crossed with Col2a1-Cre mice, miRNA transgene expression could be detected successfully by red fluorescence signals in various cartilaginous tissues. This method of creating small ncRNA transgenic mice facilitates both tissue-specific ncRNA expression and real-time visualization of its expression. It is particularly suitable forin vivostudies of the functional roles and lineage tracing of small ncRNA.
