Improved yellow-green split fluorescent proteins for protein labeling and signal amplification

Shuqin Zhou; Siyu Feng; David Brown; Bo Huang

doi:10.1101/2020.09.27.315697

Abstract

The flexibility and versatility of self-complementing split fluorescent proteins (FPs) have enabled a wide range of applications. In particular, the FP_1-10/11 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 FP_1-10/11 toolbox we previously developed, here we used a combination of directed evolution and rational design approaches, resulting in two mNeonGreen (mNG)-based split FPs (mNG3A_1-10/11 and mNG3K_1-10/11) and one mClover-based split FP (CloGFP_1-10/11). mNG3A_1-10/11 and mNG3K_1-10/11 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.

Competing Interest Statement

The authors have declared no competing interest.

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