Department of Bioengineering and Therapeutical Sciences, University of California, San Francisco, San Francisco, CA, United States of America.
School of Pharmacy, Tsinghua University, Beijing, China.
PLoS One. 2020 Nov 23;15(11):e0242592. doi: 10.1371/journal.pone.0242592. eCollection 2020.
The flexibility and versatility of self-complementing split fluorescent proteins (FPs) have enabled a wide range of applications. In particular, the FP1-10/11 split system contains a small fragment that facilitates efficient generation of endogenous-tagged cell lines and animals as well as signal amplification using tandem FP11 tags. To improve the FP1-10/11 toolbox we previously developed, here we used a combination of directed evolution and rational design approaches, resulting in two mNeonGreen (mNG)-based split FPs (mNG3A1-10/11 and mNG3K1-10/11) and one mClover-based split FP (CloGFP1-10/11). mNG3A1-10/11 and mNG3K1-10/11 not only enhanced the complementation efficiency at low expression levels, but also allowed us to demonstrate signal amplification using tandem mNG211 fragments in mammalian cells.
自互补分裂荧光蛋白 (FP) 的灵活性和多功能性使其得到了广泛的应用。特别是 FP1-10/11 分裂系统包含一个小片段，可促进高效地生成内源性标记的细胞系和动物，以及使用串联 FP11 标签进行信号放大。为了改进我们之前开发的 FP1-10/11 工具包，我们在这里使用定向进化和合理设计方法的组合，得到了两个基于 mNeonGreen (mNG) 的分裂 FP（mNG3A1-10/11 和 mNG3K1-10/11）和一个基于 mClover 的分裂 FP（CloGFP1-10/11）。mNG3A1-10/11 和 mNG3K1-10/11 不仅提高了低表达水平下的互补效率，还使我们能够在哺乳动物细胞中使用串联 mNG211 片段展示信号放大。
