RT Journal Article
SR Electronic
T1 Improved yellow-green split fluorescent proteins for protein labeling and signal amplification
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.09.27.315697
DO 10.1101/2020.09.27.315697
A1 Shuqin Zhou
A1 Siyu Feng
A1 David Brown
A1 Bo Huang
YR 2020
UL http://biorxiv.org/content/early/2020/09/28/2020.09.27.315697.abstract
AB 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.Competing Interest StatementThe authors have declared no competing interest.