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

PLoS One. 2020 Nov 23;15(11):e0242592. doi: 10.1371/journal.pone.0242592. eCollection 2020.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coloring Agents
  • Escherichia coli / metabolism
  • Fluorescent Dyes / chemistry*
  • Green Fluorescent Proteins / biosynthesis*
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Luminescent Proteins
  • Microscopy, Fluorescence / methods*

Substances

  • Coloring Agents
  • Fluorescent Dyes
  • Luminescent Proteins
  • Green Fluorescent Proteins