Two Decades of Genetically Encoded Biosensors Based on Förster Resonance Energy Transfer

Terai, Kenta; Imanishi, Ayako; Li, Chunjie

doi:10.1247/csf.18035

Cited by 43 publications

(31 citation statements)

References 112 publications

(139 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further, the enrichment of live cells can circumvent errors due to synchronization. The range of available FRET-based probes for kinase/phosphatase balances is rapidly increasing [28], and the last two decades have seen a rise in various cellular pathways studied by FRET-based reporters [29,30]. FRET-activated cell sorting (FRACS) could therefore be used to study a range of rare events.…”

Section: Discussionmentioning

confidence: 99%

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

Lafranchi

Müllers

Rutishauser

et al. 2020

Cells

View full text Add to dashboard Cite

Cells recovering from the G2/M DNA damage checkpoint rely more on Aurora A-PLK1 signaling than cells progressing through an unperturbed G2 phase, but the reason for this discrepancy is not known. Here, we devised a method based on a FRET reporter for PLK1 activity to sort cells in distinct populations within G2 phase. We employed mass spectroscopy to characterize changes in protein levels through an unperturbed G2 phase and validated that ATAD2 levels decrease in a proteasome-dependent manner. Comparing unperturbed cells with cells recovering from DNA damage, we note that at similar PLK1 activities, recovering cells contain higher levels of Cyclin B1 and increased phosphorylation of CDK1 targets. The increased Cyclin B1 levels are due to continuous Cyclin B1 production during a DNA damage response and are sustained until mitosis. Whereas partial inhibition of PLK1 suppresses mitotic entry more efficiently when cells recover from a checkpoint, partial inhibition of CDK1 suppresses mitotic entry more efficiently in unperturbed cells. Our findings provide a resource for proteome changes during G2 phase, show that the mitotic entry network is rewired during a DNA damage response, and suggest that the bottleneck for mitotic entry shifts from CDK1 to PLK1 after DNA damage.

show abstract

Section: Discussionmentioning

confidence: 99%

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

Lafranchi

Müllers

Rutishauser

et al. 2020

Cells

View full text Add to dashboard Cite

show abstract

“…Our strategy requires that barcoding proteins be spectrally separable from biosensors. A large number of fluorescent biosensors are based on detecting changes in: 1) intracellular localization or intensity of a single fluorophore, in many cases GFP; or 2) the FRET efficiency between a donor and an acceptor, most commonly cyan and yellow FPs (CFP and YFP) 8,9 . Therefore, the cyan-green-yellow range of the emission spectrum (~450-550 nm) covers a vast number of fluorescent biosensors 8 .…”

Section: Supplementary Tablementioning

confidence: 99%

Highly multiplexed imaging of biosensors in live cells

Yang

Liang

et al. 2020

Preprint

View full text Add to dashboard Cite

Fluorescent biosensors allow for real-time monitoring of biochemical activities in cells, but their multiplexing capacity is severely limited by the availability of spectral space. We overcome this problem by developing a set of barcoding proteins that are spectrally separable from commonly used FRET (fluorescence resonance energy transfer)-based and single-fluorophore biosensors. Mixed populations of barcoded cells expressing different biosensors can be concurrently imaged and computationally unmixed to achieve highly multiplexed tracking of biochemical activities in live cells.

show abstract

“…For example, the dynamic behaviors of NK cells have been demonstrated in the lymph nodes (Bajénoff et al, 2006;Beuneu et al, 2009;Mingozzi et al, 2016) and the tumor microenvironment (Barry et al, 2018;Deguine et al, 2010). Moreover, the development of genetically-encoded biosensors for signaling molecules has paved the way to monitoring of cellular activation status in not only normal but also pathological tissues (Conway et al, 2017;Terai et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Metastatic Single Tumor Cells Evade NK Cell-mediated Killing by Thrombin-mediated Loss of the Activating Ligand CD155/PVR/Necl-5

Ichise

Tsukamoto

Hirashima

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Natural killer (NK) cells lyse invading tumor cells to limit metastatic growth in the lung, but how some cancers evade this host protective mechanism to establish a growing lesion is not known. Here we have combined ultra-sensitive bioluminescence whole body imaging with intravital two-photon microscopy involving genetically-encoded biosensors to examine this question. NK cells eliminated disseminated tumor cells from the lung within 24 hrs of arrival, but not thereafter. Intravital dynamic imaging revealed that a disseminated tumor cell in a pulmonary capillary interacts with an NK cell every 2 hrs on average. In the first 4 hrs after tumor cell arrival, 50% of such encounters lead to tumor cell death but after 24 hrs of arrival, nearly 100% of the interactions result in the survival of the tumor cell. This evasion of NK cell surveillance is mediated by thrombin-dependent loss of cell surface CD155/PVR/Necl-5, a ligand for the NK cell activating receptor DNAM-1. This loss prevents the NK cell signaling needed for effective killing of tumor targets. By quantitatively visualizing the evasion of NK cell surveillance, we have uncovered a molecular mechanism for cancer evasion and provided an explanation for the anti-metastatic effect of anticoagulants.SUMMARYIntravital functional two-photon microscopy reveals that metastatic tumor cells lodged in pulmonary capillaries acquire resistance to patrolling NK cells. Protease-mediated loss of the activating ligand CD155/PVR/Necl-5 on tumor cells prevents NK cells from ERK activation and tumor cell killing.

show abstract

Two Decades of Genetically Encoded Biosensors Based on Förster Resonance Energy Transfer

Cited by 43 publications

References 112 publications

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

Highly multiplexed imaging of biosensors in live cells

Metastatic Single Tumor Cells Evade NK Cell-mediated Killing by Thrombin-mediated Loss of the Activating Ligand CD155/PVR/Necl-5

Contact Info

Product

Resources

About