The Fluorescent Protein Color Palette

Abstract

Advances in fluorescent protein development over the past 10 years have led to fine-tuning of the Aequorea victoria jellyfish color palette in the emission color range from blue to yellow, while a significant amount of progress has been achieved with reef coral species in the generation of monomeric fluorescent proteins emitting in the orange to far-red spectral regions. It is not inconceivable that near-infrared fluorescent proteins loom on the horizon. Expansion of the fluorescent protein family to include optical highlighters and FRET biosensors further arms this ubiquitous class of fluorophores with biological probes capable of photoactivation, photoconversion, and detection of molecular interactions beyond the resolution limits of optical microscopy. The success of these endeavors certainly suggests that almost any biological parameter can be investigated using the appropriate fluorescent protein-based application.

PI31 is an adaptor protein for proteasome transport in axons and required for synaptic development and function

“…Mouse proteasome subunit α4 CDS was cloned from a mouse cDNA library, and mCherry CDS was subcloned from the mCherry-Mito-7 vector (Olenych et al, 2007) (a gift from Dr. Michael Davidson, Addgene #55102). The α4-mCherry cassette was inserted between the EcoRI/NotI sites of pCAGGS/ES vector.…”

Single Molecule Fluorescence Approaches Shed Light on Intracellular RNAs

“…Especially during intracellular imaging, excited fluorophores can react with molecular oxygen within cells, resulting in the accumulation of phototoxic free radicals that can compromise sub-cellular compartments or even the entire cell's livelihood. 251 Enzymatic oxygen scavenging systems (OSS), such as those containing glucoseoxidase and catalase (GODCAT), 252 protocatechiuc acid and protocatechuate-3,4-dioxygenase (PCA/PCD) 247 or oxyfluor© 253 utilize molecular oxygen as a substrate in enzymatic reactions, thereby effectively depleting it. These OSS prevent fast photobleaching and oxygen induced free radical production, resulting in increased signal longevity.…”

Super-resolution microscopy heads towards 3D dynamics

“…Photo-chromic FPs, by contrast, are reversibly switched back and forth between a bright and a dark state driven by a cis-trans isomerization reaction. By engineered modifications of amino acids in the chromophore group or close-by regions many PS-FPs with new properties have been successfully created [86]. As PS-FPs are essentially non-fluorescent (or fluorescent at a different spectral emission band) at the start of the experiment before conversion, the number of molecules in the on-state can easily be fine-tuned by balancing the powers between the activation and imaging lasers, respectively.…”