Nonfluor Cellular Markers

Peptide antigens (epitopes) and fluorescent proteins (FPs) are the most commonly used cellular markers. To overcome the limitations of existing FP and peptide epitopes, Looger's group developed new molecular markers called 'spaghetti monster' fluorescent proteins (smFPs) that combine the advantages of both. These proteins have multiple copies of single epitope tags inserted into an FP scaffold with either an intact or a darkened chromophore. They distribute well in neurons, notably into small dendrites, spines and axons, and can be readily detected by immunolabeling. This tool set expands options for labeling and following defined populations of neurons and other cell types through brain tissue. They also perform well in preparations such as array tomography, super-resolution fluorescence imaging and electron microscopy. In living cells, the probes improve single-molecule image tracking and increase yield for RNA-seq (Viswanathan et al., 2015).

Rubin's group has generated a set of transgenes that express Flag, HA, Myc, OLLAS, or V5 tags using the darkened chromophore (aka smGdP for 'spaghetti monster Green darkened Protein') as a scaffold (Nern et al., 2015). In addition to these lines, we have another set of lines from Rubin that carry P{10xUAS-IVS-myr::smGdP-HA}attP18 and P{13xLexAop2-IVS-myr::smGdP-V5}su(Hw)attP8 in combination with different GAL4.DBDs from the split-GAL4 collection