Red fluorescent proteins (RFPs) and RFP-based biosensors for neuronal imaging applications

Shen, Yi; Lai, Tiffany; Campbell, Robert E.

doi:10.1117/1.nph.2.3.031203

Cited by 31 publications

(26 citation statements)

References 137 publications

(157 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Of particular interest are the genetically encoded optical tools based on GFP, since on the whole, these fluorescent proteins are more widely used and exhibit more predictable cellular and photophysical behavior than their red fluorescent counterparts. 42,43 We first established that BeRST 1 can perform multi-color imaging with some commonly used molecular probes for cellular organelles: the nucleus and mitochondria. We performed simultaneous three-color live cell imaging with BeRST 1 (Fig.…”

Section: Resultsmentioning

confidence: 99%

A Photostable Silicon Rhodamine Platform for Optical Voltage Sensing

2015

View full text Add to dashboard Cite

This paper describes the design and synthesis of a photostable, far-red to near-infrared (NIR) platform for optical voltage sensing. We developed a new, sulfonated silicon rhodamine fluorophore and integrated it with a phenylenevinylene molecular wire to create a Berkeley Red Sensor of Transmembrane potential, or BeRST 1 (“burst”). BeRST 1 is the first member of a class of farred to NIR voltage sensitive dyes that make use of a photoinduced electron transfer (PeT) trigger for optical interrogation of membrane voltage. We show that BeRST 1 displays bright, membrane-localized fluorescence in living cells, high photostability, and excellent voltage sensitivity in neurons. Depolarization of the plasma membrane results in rapid fluorescence increases (24% ΔF/F per 100 mV). BeRST 1 can be used in conjunction with fluorescent stains for organelles, Ca2+ indicators, and voltage-sensitive fluorescent proteins. In addition, the red-shifted spectral profile of BeRST 1, relative to commonly employed optogenetic actuators like ChannelRhodopsin2 (ChR2), which require blue light, enables optical electrophysiology in neurons. The high speed, sensitivity, photostability and long-wavelength fluorescence profiles of BeRST 1 make it a useful platform for the non-invasive, optical dissection of neuronal activity.

show abstract

Section: Resultsmentioning

confidence: 99%

A Photostable Silicon Rhodamine Platform for Optical Voltage Sensing

2015

View full text Add to dashboard Cite

show abstract

“…A major conceptual innovation in ratiometric sensor design known as “dimerization-dependent fluorescent protein exchange” or “FPX” [29, 30] was recently devised by Robert Campbell’s laboratory. This is a somewhat complex strategy that, for the case of the FPX cAMP reporter (hPHT-PKA), requires cotransfection with three different constructs.…”

Section: Sensors Based On Native Mammalian Effectors For Camp: Pkamentioning

confidence: 99%

Interrogating cyclic AMP signaling using optical approaches

et al. 2017

View full text Add to dashboard Cite

Optical reporters for cAMP represent a fundamental advancement in our ability to investigate the dynamics of cAMP signaling. These fluorescent sensors can measure changes in cAMP in single cells or in microdomains within cells as opposed to whole populations of cells required for other methods of measuring cAMP. The first optical cAMP reporters were FRET-based sensors utilizing dissociation of purified regulatory and catalytic subunits of PKA, introduced by Roger Tsien in the early 1990s. The utility of these sensors was vastly improved by creating genetically encoded versions that could be introduced into cells with transfection, the first of which was published in the year 2000. Subsequently, improved sensors have been developed using different cAMP binding platforms, optimized fluorescent proteins, and targeting motifs that localize to specific microdomains. The most common sensors in use today are FRET-based sensors designed around an Epac backbone. These rely on the significant conformational changes in Epac when it binds cAMP, altering the signal between FRET pairs flanking Epac. Several other strategies for optically interrogating cAMP have been developed, including fluorescent translocation reporters, dimerization-dependent FP based biosensors, BRET (bioluminescence resonance energy transfer)-based sensors, non-FRET single wavelength reporters, and sensors based on bacterial cAMP-binding domains. Other newly described mammalian cAMP-binding proteins such as Popdc and CRIS may someday be exploited in sensor design. With the proliferation of engineered fluorescent proteins and the abundance of cAMP binding targets in nature, the field of optical reporters for cAMP should continue to see rapid refinement in the coming years.

show abstract

“…Engineering of eqFP611 produced mRuby, mRuby2, and mRuby3, a line of RFPs with relatively large stokes-shift and bright red fluorescence [10][11][12] . Together, these three lineages of monomeric RFPs are commonly used in a variety of fluorescence imaging applications and have served as templates for developing red fluorescent indicators of various biochemical activities 13 .…”

Section: Introductionmentioning

confidence: 99%

“…Overall, K-GECO1 unravels robust responses to cytosolic Ca 2+ concentration changes in neurons in organotypic brain slices. 13 In vivo Ca 2+ imaging with K-GECO1. To test K-GECO1 in zebrafish spinal cord sensory neurons in vivo, we transiently expressed K-GECO1 in Rohon Beard cells.…”

Section: Introductionmentioning

confidence: 99%