The Growing and Glowing Toolbox of Fluorescent and Photoactive Proteins.
Over the past 20 years, protein engineering has been extensively used to improve and modify the fundamental properties of fluorescent proteins (FPs) with the goal of adapting them for a fantastic range of applications. FPs have been modified by a combination of rational design, structure-based mutagenesis, and countless cycles of directed evolution (gene diversification followed by selection of clones with desired properties) that have collectively pushed the properties to photophysical and biochemical extremes. In this review, we provide both a summary of the progress that has been made during the past two decades, and a broad overview of the current state of FP development and applications in mammalian systems.
Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: ear001@ucsd.edu.
Erik A
EA
Rodriguez
Department of Chemistry, University of Alberta, Edmonton, AB, T6G 2G2, Canada. Electronic address: rc4@ualberta.ca.
Robert E
RE
Campbell
School of Medicine, University of Tasmania, Hobart, TAS 7000, Australia. Electronic address: john.lin@utas.edu.au.
John Y
JY
Lin
Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA; Department of Pediatrics, Stanford University, Stanford, CA, 94305, USA. Electronic address: mzlin@stanford.edu.
Michael Z
MZ
Lin
Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan. Electronic address: matsushi@brain.riken.go.jp.
Atsushi
A
Miyawaki
Department of Chemistry and Biochemistry, BioFrontiers Institute, University of Colorado, Boulder, CO, 80303, USA. Electronic address: amy.palmer@colorado.edu.
Amy E
AE
Palmer
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, 94158, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA, 94158, USA. Electronic address: xiaokun.shu@ucsf.edu.
Xiaokun
X
Shu
Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: jzhang32@ucsd.edu.
Jin
J
Zhang
Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA; Howard Hughes Medical Institute, University of California, San Diego, La Jolla, CA, 92093, USA. Electronic address: rtsien@ucsd.edu.
Roger Y
RY
Tsien
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics*; D010834:Phytochrome / Q000737:chemistry; D015202:Protein Engineering
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010834:Phytochrome