Development and Applications of Superfolder and Split Fluorescent Protein Detection Systems in Biology

• Published in: International journal of molecular sciences Vol. 20; no. 14; p. 3479
• Main Authors: Pedelacq, Jean-Denis, Cabantous, Stéphanie
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 15.07.2019; MDPI
• Subjects: Animals; Biochemistry, Molecular Biology; Biotechnology; E coli; Energy transfer; Engineering; Evolution; Experiments; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Green Fluorescent Proteins - standards; Humans; Life Sciences; Localization; Microscopy, Fluorescence - methods; Mutagenesis; Mutation; Neurosciences; Protein Engineering - methods; Protein Folding; Proteins; Review; Sensors; fluorescent protein; tripartite; folding; PPI; split-GFP; bipartite; superfolder
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms20143479

Molecular engineering of the green fluorescent protein (GFP) into a robust and stable variant named Superfolder GFP (sfGFP) has revolutionized the field of biosensor development and the use of fluorescent markers in diverse area of biology. sfGFP-based self-associating bipartite split-FP systems have been widely exploited to monitor soluble expression in vitro, localization, and trafficking of proteins in cellulo. A more recent class of split-FP variants, named « tripartite » split-FP, that rely on the self-assembly of three GFP fragments, is particularly well suited for the detection of protein–protein interactions. In this review, we describe the different steps and evolutions that have led to the diversification of superfolder and split-FP reporter systems, and we report an update of their applications in various areas of biology, from structural biology to cell biology.