Open AccessImproved yellow-green split fluorescent proteins for protein labeling and signal amplification
Author(s) -
Shuqin Zhou,
Siyu Feng,
David Brown,
Bo Huang
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0242592
Subject(s) - green fluorescent protein , fluorescence , tandem , bimolecular fluorescence complementation , computational biology , protein engineering , signal peptide , chemistry , biology , peptide sequence , genetics , biochemistry , physics , enzyme , materials science , gene , quantum mechanics , composite material
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 FP 11 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 mNG2 11 fragments in mammalian cells.