Application of fluorescent proteins and RCAS-vector in cell lineage analysis

• Main Authors: Hsin-Ying Lin, 林欣穎
• Other Authors: 鄭旭辰
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/83321277519872975659

碩士 === 國立中興大學 === 生命科學系所 === 97 === Cells either migrate as individuals or as clusters to their destinations. To record the cell lineages, it will be necessary to label these cells and track their migratory processes. Fluorescence proteins or fluorescence dyes are commonly used for labeling cells for live imaging. However, the labeling was limited to one or two colors for each sample. Recent improvement of imaging technology allowed compound labeling of multiple fluorophores(fluorochrome) on different cells can be distinguished and identified separately, such as the Brainbow transgenic mice. They employed Cre/lox recombination to create a stochastic choice of expression between three or more fluorescent proteins (XFPs). The differential expression of XFP mixtures generated a population of labeled cells with up to 90 distinguishable colors. Avian replication-competent retroviruses (RCAS) allow efficient infection of proliferating cells and stable integration into the host chromosome. A Proliferating cell can be infected only one RCAS retrovirus, and the cell couldn’t be infected other retrovirus. We used RCAS viral vectors that carry different fluorescent protein cDNAs to simultaneously infect proliferating cells that would create many cells with individually identifiable colors for tracking cell lineages and cell migration. Therefore, avian RCAS-XFP may serve as a powerful tool to multicolor staining of single cell and to study cell migration mechanisms underlying avian development.