Issue 32, 2023

Tri-part NanoLuc as a new split technology with potential applications in chemical biology: a mini-review

Abstract

For several decades, researchers have been using protein-fragment complementation assay (PCA) approaches for biosensing to study protein–protein interaction for a variety of aims, including viral infection, cellular apoptosis, G protein coupled receptor (GPCR) signaling, drug and substrate screening, and protein aggregation and protein editing by CRISPR/Cas9. As a reporter, NanoLuc (NLuc), a smaller and the brightest engineered luciferase derived from deep-sea shrimp Oplophorus gracilirostris, has been found to have many benefits over other luminescent enzymes in PCA. Inspired by the split green fluorescent protein (GFP) and its β-barrel structure, two split NLuc consisting of peptide fragments have been reported including the binary and ternary NLuc systems. NanoBiT® (large fragment + peptide) has been used extensively. In contrast, tripart split NLuc (large fragment + 2 peptides) has been applied and hardly used, while it has some advantages over NanoBiT in some studies. Nevertheless, tripart NLuc has some drawbacks and challenges to overcome but has several potential characteristics to become a multifunctional and powerful tool. In this review, several aspects of tripart NLuc are studied and a brief comparison with NanoBiT® is given.

Graphical abstract: Tri-part NanoLuc as a new split technology with potential applications in chemical biology: a mini-review

Article information

Article type
Minireview
Submitted
05 Apr 2023
Accepted
10 Jul 2023
First published
10 Jul 2023

Anal. Methods, 2023,15, 3924-3931

Tri-part NanoLuc as a new split technology with potential applications in chemical biology: a mini-review

M. Oliayi, R. Emamzadeh, M. Rastegar and M. Nazari, Anal. Methods, 2023, 15, 3924 DOI: 10.1039/D3AY00512G

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