![]() ![]() However, for the long 30 years since the beginning of this story more than 45 years ago ( 392), green fluorescent protein (GFP) was of interest only to a handful of scientists studying luminescence of marine creatures. The development of green pigs and cats has been well covered by the media, and transgenic fluorescent fish are common in home aquaria. Here we focus on the structure, evolution, and function of GFP-like proteins and their numerous applications for in vivo imaging, with particular attention to recent techniques.Ĭurrently, the term green fluorescent protein is well known beyond the realm of life science. ![]() Fast-maturing fluorescent proteins, cell clocks, and timers further expand the options for real time studies in living tissues. ![]() Genetically encoded sensors make it possible to monitor the activity of enzymes and the concentrations of various analytes. Photoactivatable fluorescent proteins enable tracking of photolabeled molecules and cells in space and time and can also be used for super-resolution imaging. The diversity of currently available fluorescent proteins covers nearly the entire visible spectrum, providing numerous alternative possibilities for multicolor labeling and studies of protein interactions. Many laboratories have focused their efforts on identification and development of fluorescent proteins with novel characteristics and enhanced properties, resulting in a powerful toolkit for visualization of structural organization and dynamic processes in living cells and organisms. Our integrated STARTRAC analyses provide a powerful approach to dissect the T cell properties in colorectal cancer comprehensively, and could provide insights into the dynamic relationships of T cells in other cancers.Green fluorescent protein (GFP) from the jellyfish Aequorea victoria and its homologs from diverse marine animals are widely used as universal genetically encoded fluorescent labels. Furthermore, IGFLR1 was highly expressed in both CXCL13 +BHLHE40 + T H1-like cells and CD8 + exhausted T cells and possessed co-stimulatory functions. Only CXCL13 +BHLHE40 + T H1-like cells were preferentially enriched in patients with microsatellite-instable tumours, and this might explain their favourable responses to immune-checkpoint blockade. Notably, we identified two IFNG + T H1-like cell clusters in tumours that were associated with distinct IFNγ-regulating transcription factors -the GZMK + effector memory T cells, which were associated with EOMES and RUNX3, and CXCL13 +BHLHE40 + T H1-like cell clusters, which were associated with BHLHE40. Of the CD4 + T cells, most tumour-infiltrating T regulatory (T reg) cells showed clonal exclusivity, whereas certain T reg cell clones were developmentally linked to several T helper (T H) cell clones. Although both CD8 + effector and 'exhausted' T cells exhibited high clonal expansion, they were independently connected with tumour-resident CD8 + effector memory cells, implicating a TCR-based fate decision. Here we obtained transcriptomes of 11,138 single T cells from 12 patients with colorectal cancer, and developed single T cell analysis by RNA sequencing and TCR tracking (STARTRAC) indices to quantitatively analyse the dynamic relationships among 20 identified T cell subsets with distinct functions and clonalities. The enormous T cell receptor (TCR) repertoire, which is required for the recognition of foreign and self-antigens 2, could serve as lineage tags to track these T cells in tumours 3. T cells are key elements of cancer immunotherapy 1 but certain fundamental properties, such as the development and migration of T cells within tumours, remain unknown. ![]()
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