Improved yellow-green split fluorescent proteins for protein labeling and signal amplification.
basic_science · Level V
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- Record sourced from PubMed, PMID 33227014.
- Also identified by DOI 10.1371/journal.pone.0242592 and PMC identifier 7682878.
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Abstract
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.
Medical subject headings
- Fluorescent Dyes
- Green Fluorescent Proteins
- Microscopy, Fluorescence