Dynamic FRET-FLIM based screening of signal transduction pathways

Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attracti...

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Veröffentlicht in:	Scientific reports Jg. 11; H. 1; S. 20711 - 13
Hauptverfasser:	Harkes, Rolf, Kukk, Olga, Mukherjee, Sravasti, Klarenbeek, Jeffrey, van den Broek, Bram, Jalink, Kees
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	London Nature Publishing Group UK 20.10.2021 Nature Publishing Group Nature Portfolio
Schlagworte:	631/114/1314 631/114/2163 631/114/2391 631/114/2400 631/45/731 631/57/2267 631/61/191/505 631/80/2373 631/80/86 Cell activation Cell Line, Tumor Cell signaling Cell surface Cyclic AMP Cyclic AMP - metabolism Energy transfer Fluorescence Fluorescence resonance energy transfer Fluorescence Resonance Energy Transfer - methods HeLa Cells Humanities and Social Sciences Humans Instrumentation Isoforms Microscopy, Fluorescence - methods multidisciplinary Optical Imaging - methods Phosphoric Diester Hydrolases - metabolism Photons Protein interaction Proteins - metabolism Science Science (multidisciplinary) Signal transduction Signal Transduction - physiology
ISSN:	2045-2322, 2045-2322
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Abstract	Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-S H189 . Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput.
AbstractList	Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-SH189. Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput. Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-S H189 . Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput. Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein-protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-S . Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput. Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein-protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-SH189. Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput.Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein-protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-SH189. Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput. Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-SH189. Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput. Abstract Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the outcome of signal transduction events. With new highly sensitive and photon efficient FLIM instrumentation, the technique also becomes attractive to screen, with high temporal resolution, for fast changes in Förster Resonance Energy Transfer (FRET), such as those occurring upon activation of cell signaling. The second messenger cyclic adenosine monophosphate (cAMP) is rapidly formed following activation of certain cell surface receptors. cAMP is subsequently degraded by a set of phosphodiesterases (PDEs) which display cell-type specific expression and may also affect baseline levels of the messenger. To study which specific PDEs contribute most to cAMP regulation, we knocked down individual PDEs and recorded breakdown rates of cAMP levels following transient stimulation in HeLa cells stably expressing the FRET/FLIM sensor, Epac-SH189. Many hundreds of cells were recorded at 5 s intervals for each condition. FLIM time traces were calculated for every cell, and decay kinetics were obtained. cAMP clearance was significantly slower when PDE3A and, to a lesser amount, PDE10A were knocked down, identifying these isoforms as dominant in HeLa cells. However, taking advantage of the quantitative FLIM data, we found that knockdown of individual PDEs has a very limited effect on baseline cAMP levels. By combining photon-efficient FLIM instrumentation with optimized sensors, systematic gene knockdown and an automated open-source analysis pipeline, our study demonstrates that dynamic screening of transient cell signals has become feasible. The quantitative platform described here provides detailed kinetic analysis of cellular signals in individual cells with unprecedented throughput.
ArticleNumber	20711
Author	Jalink, Kees Harkes, Rolf Kukk, Olga Mukherjee, Sravasti van den Broek, Bram Klarenbeek, Jeffrey
Author_xml	– sequence: 1 givenname: Rolf surname: Harkes fullname: Harkes, Rolf organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute – sequence: 2 givenname: Olga surname: Kukk fullname: Kukk, Olga organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute – sequence: 3 givenname: Sravasti surname: Mukherjee fullname: Mukherjee, Sravasti organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute, Swammerdam Institute for Life Sciences, University of Amsterdam – sequence: 4 givenname: Jeffrey surname: Klarenbeek fullname: Klarenbeek, Jeffrey organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute – sequence: 5 givenname: Bram surname: van den Broek fullname: van den Broek, Bram organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute, BioImaging Facility, The Netherlands Cancer Institute – sequence: 6 givenname: Kees surname: Jalink fullname: Jalink, Kees email: k.jalink@nki.nl organization: Cell Biophysics Group, Department of Cell Biology, The Netherlands Cancer Institute, Swammerdam Institute for Life Sciences, University of Amsterdam
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CitedBy_id	crossref_primary_10_1016_j_cell_2022_07_026 crossref_primary_10_3390_bios13100939 crossref_primary_10_1038_s41598_023_30098_w crossref_primary_10_3390_bios15090624 crossref_primary_10_1002_1873_3468_70055 crossref_primary_10_1134_S1990519X22050078 crossref_primary_10_1111_bph_16297 crossref_primary_10_1002_1873_3468_15093 crossref_primary_10_3390_cells13131114
Cites_doi	10.1038/nmeth.3836 10.1016/j.ceb.2013.11.009 10.1146/annurev.biochem.76.060305.150444 10.1016/j.cellsig.2018.03.002 10.1074/mcp.T700006-MCP200 10.1529/biophysj.108.131276 10.1371/journal.pone.0019170 10.1038/srep28186 10.1016/S0955-0674(99)00073-3 10.1111/fcp.12080 10.1007/s00424-017-2106-8 10.1073/pnas.1903808116 10.1371/journal.pone.0047826 10.1074/jbc.M202753200 10.1038/sj.tpj.6500393 10.1016/j.coph.2011.10.006 10.1038/s41592-019-0618-1 10.1016/j.coph.2011.09.001 10.1016/j.cell.2020.07.043 10.1093/cvr/cvy161 10.7554/eLife.58039 10.1038/nmeth.4196 10.1016/S0079-6603(00)66031-2 10.1371/journal.pone.0122513 10.1083/jcb.200605050 10.1038/ncomms15031 10.1038/ng.3302 10.1016/j.tibs.2009.09.007 10.1038/nbt.3536 10.1074/jbc.M706765200 10.1074/jbc.272.11.6823 10.3109/10799893.2010.509728 10.1038/s41598-020-61478-1 10.1124/pr.113.008219 10.1038/s41573-019-0033-4 10.1038/nrd4228 10.1016/j.cell.2015.11.007 10.1016/j.cell.2019.09.016 10.1016/j.tips.2017.11.006 10.1016/j.tips.2017.11.009 10.1016/j.cell.2020.07.035 10.1111/brv.12020 10.1038/s41592-020-01018-x 10.1186/gb-2007-8-s1-s7 10.1073/pnas.1806318115 10.1016/S0075-7535(08)00007-7
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References	Margineanu (CR9) 2016; 6 Begum, Shen, Manganiello (CR45) 2011; 11 Zhang (CR18) 2020; 182 Klarenbeek, Goedhart, Hink, Gadella, Jalink (CR24) 2011; 6 Gadella (CR6) 2011 Baillie, Tejeda, Kelly (CR46) 2019; 18 Bassler, Schultz, Lupas (CR14) 2018; 46 Wang, Lu, Emanuel, Babcock, Zhuang (CR3) 2019; 116 Bock (CR49) 2020; 182 Eichel, von Zastrow (CR17) 2018; 39 Gancedo (CR13) 2013; 88 Lohse, Maiellaro, Calebiro (CR33) 2014; 27 Willoughby (CR23) 2007; 282 Zhai (CR42) 2012; 7 Maass (CR44) 2015; 47 Maurice (CR22) 2014; 13 Burdyga (CR50) 2018; 115 CR5 Klarenbeek, Goedhart, van Batenburg, Groenewald, Jalink (CR25) 2015; 10 Levitt (CR11) 2020; 10 Boutros, Heigwer, Laufer (CR1) 2015; 163 Soderling, Beavo (CR31) 2000; 12 Evers (CR30) 2016; 34 Otero (CR20) 2014; 28 Houslay (CR38) 2010; 35 Degerman, Belfrage, Manganiello (CR40) 1997; 272 Lazar (CR35) 2020; 9 Feldman (CR4) 2019; 179 Idres (CR43) 2019; 115 Musheshe, Schmidt, Zaccolo (CR15) 2018; 39 Alvarez, Widzgowski, Ossato, van den Broek, Jalink, Kuschel, Roberti, Hecht (CR10) 2019; 16 Dünnes (CR41) 2018; 470 Esposito, Dohm, Bähr, Wouters (CR7) 2007; 6 Terrin (CR48) 2006; 175 Ahles, Engelhardt (CR37) 2014; 66 Zamah, Delahunty, Luttrell, Lefkowitz (CR39) 2002; 277 Movsesian, Wever-Pinzon, Vandeput (CR47) 2011; 11 CR29 CR28 Shakur (CR32) 2001; 66 CR21 Vilardaga (CR34) 2010; 30 Taylor (CR36) 2007; 7 Padilla-Parra, Audugé, Coppey-Moisan, Tramier (CR8) 2008; 95 Stringer, Wang, Michaelos, Pachitariu (CR26) 2021; 18 Surdo (CR16) 2017; 8 Wagner, Klein (CR2) 2017; 14 Raspe (CR12) 2016; 13 Kawakami (CR27) 2007; 8 Conti, Beavo (CR19) 2007; 76 A Bock (98_CR49) 2020; 182 D Feldman (98_CR4) 2019; 179 98_CR5 TWJ Gadella (98_CR6) 2011 M Conti (98_CR19) 2007; 76 M Movsesian (98_CR47) 2011; 11 JB Klarenbeek (98_CR24) 2011; 6 C Wang (98_CR3) 2019; 116 DH Maurice (98_CR22) 2014; 13 B Evers (98_CR30) 2016; 34 A Burdyga (98_CR50) 2018; 115 D Willoughby (98_CR23) 2007; 282 C Stringer (98_CR26) 2021; 18 N Begum (98_CR45) 2011; 11 E Degerman (98_CR40) 1997; 272 A Margineanu (98_CR9) 2016; 6 NC Surdo (98_CR16) 2017; 8 AM Lazar (98_CR35) 2020; 9 98_CR21 JA Levitt (98_CR11) 2020; 10 C Otero (98_CR20) 2014; 28 J Bassler (98_CR14) 2018; 46 AM Zamah (98_CR39) 2002; 277 MRG Taylor (98_CR36) 2007; 7 DE Wagner (98_CR2) 2017; 14 K Eichel (98_CR17) 2018; 39 Y Shakur (98_CR32) 2001; 66 98_CR28 M Boutros (98_CR1) 2015; 163 98_CR29 JZ Zhang (98_CR18) 2020; 182 S Padilla-Parra (98_CR8) 2008; 95 K Kawakami (98_CR27) 2007; 8 PG Maass (98_CR44) 2015; 47 S Idres (98_CR43) 2019; 115 M Raspe (98_CR12) 2016; 13 MD Houslay (98_CR38) 2010; 35 A Esposito (98_CR7) 2007; 6 S Dünnes (98_CR41) 2018; 470 A Ahles (98_CR37) 2014; 66 J-P Vilardaga (98_CR34) 2010; 30 J Klarenbeek (98_CR25) 2015; 10 SH Soderling (98_CR31) 2000; 12 JM Gancedo (98_CR13) 2013; 88 MJ Lohse (98_CR33) 2014; 27 LAJ Alvarez (98_CR10) 2019; 16 K Zhai (98_CR42) 2012; 7 A Terrin (98_CR48) 2006; 175 N Musheshe (98_CR15) 2018; 39 GS Baillie (98_CR46) 2019; 18
References_xml	– volume: 13 start-page: 501 year: 2016 end-page: 504 ident: CR12 article-title: siFLIM: Single-image frequency-domain FLIM provides fast and photon-efficient lifetime data publication-title: Nat. Methods doi: 10.1038/nmeth.3836 – volume: 27 start-page: 87 year: 2014 end-page: 93 ident: CR33 article-title: Kinetics and mechanism of G protein-coupled receptor activation publication-title: Curr. Opin. Cell Biol. doi: 10.1016/j.ceb.2013.11.009 – volume: 76 start-page: 481 year: 2007 end-page: 511 ident: CR19 article-title: Biochemistry and physiology of cyclic nucleotide phosphodiesterases: Essential components in cyclic nucleotide signaling publication-title: Annu. Rev. Biochem. doi: 10.1146/annurev.biochem.76.060305.150444 – volume: 46 start-page: 135 year: 2018 end-page: 144 ident: CR14 article-title: Adenylate cyclases: Receivers, transducers, and generators of signals publication-title: Cell. Signal. doi: 10.1016/j.cellsig.2018.03.002 – volume: 6 start-page: 1446 year: 2007 end-page: 1454 ident: CR7 article-title: Unsupervised fluorescence lifetime imaging microscopy for high content and high throughput screening publication-title: Mol. Cell. Proteomics doi: 10.1074/mcp.T700006-MCP200 – volume: 95 start-page: 2976 year: 2008 end-page: 2988 ident: CR8 article-title: Quantitative FRET analysis by fast acquisition time domain FLIM at high spatial resolution in living cells publication-title: Biophys. J. doi: 10.1529/biophysj.108.131276 – volume: 6 start-page: e19170 year: 2011 ident: CR24 article-title: A mTurquoise-based cAMP sensor for both FLIM and ratiometric read-out has improved dynamic range publication-title: PLoS ONE doi: 10.1371/journal.pone.0019170 – ident: CR29 – volume: 6 start-page: 28186 year: 2016 ident: CR9 article-title: Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) publication-title: Sci. Rep. doi: 10.1038/srep28186 – volume: 12 start-page: 174 year: 2000 end-page: 179 ident: CR31 article-title: Regulation of cAMP and cGMP signaling: New phosphodiesterases and new functions publication-title: Curr. Opin. Cell Biol. doi: 10.1016/S0955-0674(99)00073-3 – volume: 28 start-page: 593 year: 2014 end-page: 607 ident: CR20 article-title: Temporal and spatial regulation of cAMP signaling in disease: Role of cyclic nucleotide phosphodiesterases publication-title: Fundam. Clin. Pharmacol. doi: 10.1111/fcp.12080 – volume: 470 start-page: 693 year: 2018 end-page: 702 ident: CR41 article-title: Phosphodiesterase 3A expression and activity in the murine vasculature is influenced by NO-sensitive guanylyl cyclase publication-title: Pflüg. Arch. Eur. J. Physiol. doi: 10.1007/s00424-017-2106-8 – ident: CR21 – volume: 116 start-page: 10842 year: 2019 end-page: 10851 ident: CR3 article-title: Imaging-based pooled CRISPR screening reveals regulators of lncRNA localization publication-title: Proc. Natl. Acad. Sci. doi: 10.1073/pnas.1903808116 – volume: 7 start-page: e47826 year: 2012 ident: CR42 article-title: β-Adrenergic cAMP signals are predominantly regulated by phosphodiesterase type 4 in cultured adult rat aortic smooth muscle cells publication-title: PLoS ONE doi: 10.1371/journal.pone.0047826 – volume: 277 start-page: 31249 year: 2002 end-page: 31256 ident: CR39 article-title: Protein Kinase A-mediated phosphorylation of the β2-adrenergic receptor regulates its coupling to Gs and Gi demonstration in a reconstituted systeM publication-title: J. Biol. Chem. doi: 10.1074/jbc.M202753200 – volume: 7 start-page: 29 year: 2007 end-page: 37 ident: CR36 article-title: Pharmacogenetics of the human beta-adrenergic receptors publication-title: Pharmacogenom. J. doi: 10.1038/sj.tpj.6500393 – volume: 11 start-page: 725 year: 2011 end-page: 729 ident: CR45 article-title: Role of PDE3A in regulation of cell cycle progression in mouse vascular smooth muscle cells and oocytes: Implications in cardiovascular diseases and infertility publication-title: Curr. Opin. Pharmacol. doi: 10.1016/j.coph.2011.10.006 – volume: 16 start-page: 1069 year: 2019 end-page: 1071 ident: CR10 article-title: SP8 FALCON: A novel concept in fluorescence lifetime imaging enabling video-rate confocal FLIM publication-title: Nat. Methods doi: 10.1038/s41592-019-0618-1 – volume: 11 start-page: 707 year: 2011 end-page: 713 ident: CR47 article-title: PDE3 inhibition in dilated cardiomyopathy publication-title: Curr. Opin. Pharmacol. doi: 10.1016/j.coph.2011.09.001 – volume: 182 start-page: 1531 year: 2020 end-page: 1544.e15 ident: CR18 article-title: Phase separation of a PKA regulatory subunit controls cAMP compartmentation and oncogenic signaling publication-title: Cell doi: 10.1016/j.cell.2020.07.043 – volume: 115 start-page: 130 year: 2019 end-page: 144 ident: CR43 article-title: Contribution of BKCa channels to vascular tone regulation by PDE3 and PDE4 is lost in heart failure publication-title: Cardiovasc. Res. doi: 10.1093/cvr/cvy161 – ident: CR5 – volume: 9 start-page: e58039 year: 2020 ident: CR35 article-title: G protein-regulated endocytic trafficking of adenylyl cyclase type 9 publication-title: Elife doi: 10.7554/eLife.58039 – volume: 14 start-page: 237 year: 2017 end-page: 238 ident: CR2 article-title: Genetic screening enters the single-cell era publication-title: Nat. Methods doi: 10.1038/nmeth.4196 – volume: 66 start-page: 241 year: 2001 end-page: 277 ident: CR32 article-title: Regulation and function of the cyclic nucleotide phosphodiesterase (PDE3) gene family publication-title: Prog. Nucleic Acid Res. Mol. Biol. doi: 10.1016/S0079-6603(00)66031-2 – volume: 10 start-page: e0122513 year: 2015 ident: CR25 article-title: Fourth-generation Epac-based FRET sensors for cAMP feature exceptional brightness, photostability and dynamic range: Characterization of dedicated sensors for FLIM, for ratiometry and with high affinity publication-title: PLoS ONE doi: 10.1371/journal.pone.0122513 – volume: 175 start-page: 441 year: 2006 end-page: 451 ident: CR48 article-title: PGE1 stimulation of HEK293 cells generates multiple contiguous domains with different [cAMP]: Role of compartmentalized phosphodiesterases publication-title: J. Cell Biol. doi: 10.1083/jcb.200605050 – volume: 8 start-page: 15031 year: 2017 ident: CR16 article-title: FRET biosensor uncovers cAMP nano-domains at β-adrenergic targets that dictate precise tuning of cardiac contractility publication-title: Nat. Commun. doi: 10.1038/ncomms15031 – volume: 47 start-page: 647 year: 2015 end-page: 653 ident: CR44 article-title: PDE3A mutations cause autosomal dominant hypertension with brachydactyly publication-title: Nat. Genet. doi: 10.1038/ng.3302 – year: 2011 ident: CR6 publication-title: FRET and FLIM Techniques – volume: 35 start-page: 91 year: 2010 end-page: 100 ident: CR38 article-title: Underpinning compartmentalised cAMP signalling through targeted cAMP breakdown publication-title: Trends Biochem. Sci. doi: 10.1016/j.tibs.2009.09.007 – volume: 34 start-page: 631 year: 2016 end-page: 633 ident: CR30 article-title: CRISPR knockout screening outperforms shRNA and CRISPRi in identifying essential genes publication-title: Nat. Biotechnol. doi: 10.1038/nbt.3536 – volume: 282 start-page: 34235 year: 2007 end-page: 34249 ident: CR23 article-title: Dynamic regulation, desensitization, and cross-talk in discrete subcellular microdomains during β2-adrenoceptor and prostanoid receptor cAMP signaling publication-title: J. Biol. Chem. doi: 10.1074/jbc.M706765200 – volume: 272 start-page: 6823 year: 1997 end-page: 6826 ident: CR40 article-title: Structure, localization, and regulation of cGMP-inhibited phosphodiesterase (PDE3) publication-title: J. Biol. Chem. doi: 10.1074/jbc.272.11.6823 – volume: 30 start-page: 304 year: 2010 end-page: 312 ident: CR34 article-title: Theme and variations on kinetics of GPCR activation/deactivation publication-title: J. Recept. Signal Transduct. Res. doi: 10.3109/10799893.2010.509728 – volume: 10 start-page: 5146 year: 2020 ident: CR11 article-title: Quantitative real-time imaging of intracellular FRET biosensor dynamics using rapid multi-beam confocal FLIM publication-title: Sci. Rep. doi: 10.1038/s41598-020-61478-1 – volume: 66 start-page: 598 year: 2014 end-page: 637 ident: CR37 article-title: Polymorphic variants of adrenoceptors: Pharmacology, physiology, and role in disease publication-title: Pharmacol. Rev. doi: 10.1124/pr.113.008219 – volume: 18 start-page: 770 year: 2019 end-page: 796 ident: CR46 article-title: Therapeutic targeting of 3′,5′-cyclic nucleotide phosphodiesterases: inhibition and beyond publication-title: Nat. Rev. Drug Discov. doi: 10.1038/s41573-019-0033-4 – volume: 13 start-page: 290 year: 2014 end-page: 314 ident: CR22 article-title: Advances in targeting cyclic nucleotide phosphodiesterases publication-title: Nat. Rev. Drug Discov. doi: 10.1038/nrd4228 – volume: 163 start-page: 1314 year: 2015 end-page: 1325 ident: CR1 article-title: Microscopy-based high-content screening publication-title: Cell doi: 10.1016/j.cell.2015.11.007 – volume: 179 start-page: 787 year: 2019 end-page: 799.e17 ident: CR4 article-title: Optical pooled screens in human cells publication-title: Cell doi: 10.1016/j.cell.2019.09.016 – volume: 39 start-page: 209 year: 2018 end-page: 222 ident: CR15 article-title: cAMP: From long-range second messenger to nanodomain signalling publication-title: Trends Pharmacol. Sci. doi: 10.1016/j.tips.2017.11.006 – volume: 39 start-page: 200 year: 2018 end-page: 208 ident: CR17 article-title: Subcellular organization of GPCR signaling publication-title: Trends Pharmacol. Sci. doi: 10.1016/j.tips.2017.11.009 – volume: 182 start-page: 1519 year: 2020 end-page: 1530.e17 ident: CR49 article-title: Optical mapping of cAMP signaling at the nanometer scale publication-title: Cell doi: 10.1016/j.cell.2020.07.035 – volume: 88 start-page: 645 year: 2013 end-page: 668 ident: CR13 article-title: Biological roles of cAMP: Variations on a theme in the different kingdoms of life publication-title: Biol. Rev. Camb. Philos. Soc. doi: 10.1111/brv.12020 – volume: 18 start-page: 100 year: 2021 end-page: 106 ident: CR26 article-title: Cellpose: A generalist algorithm for cellular segmentation publication-title: Nat. Methods doi: 10.1038/s41592-020-01018-x – volume: 8 start-page: S7 year: 2007 ident: CR27 article-title: Tol2: A versatile gene transfer vector in vertebrates publication-title: Genome Biol. doi: 10.1186/gb-2007-8-s1-s7 – ident: CR28 – volume: 115 start-page: E6497 year: 2018 end-page: E6506 ident: CR50 article-title: Phosphatases control PKA-dependent functional microdomains at the outer mitochondrial membrane publication-title: Proc. Natl. Acad. Sci. doi: 10.1073/pnas.1806318115 – volume: 16 start-page: 1069 year: 2019 ident: 98_CR10 publication-title: Nat. Methods doi: 10.1038/s41592-019-0618-1 – volume: 115 start-page: E6497 year: 2018 ident: 98_CR50 publication-title: Proc. Natl. Acad. Sci. doi: 10.1073/pnas.1806318115 – volume: 6 start-page: 28186 year: 2016 ident: 98_CR9 publication-title: Sci. Rep. doi: 10.1038/srep28186 – volume: 35 start-page: 91 year: 2010 ident: 98_CR38 publication-title: Trends Biochem. Sci. doi: 10.1016/j.tibs.2009.09.007 – volume: 6 start-page: 1446 year: 2007 ident: 98_CR7 publication-title: Mol. Cell. Proteomics doi: 10.1074/mcp.T700006-MCP200 – ident: 98_CR28 – volume: 27 start-page: 87 year: 2014 ident: 98_CR33 publication-title: Curr. Opin. Cell Biol. doi: 10.1016/j.ceb.2013.11.009 – volume: 10 start-page: e0122513 year: 2015 ident: 98_CR25 publication-title: PLoS ONE doi: 10.1371/journal.pone.0122513 – ident: 98_CR5 doi: 10.1016/S0075-7535(08)00007-7 – volume: 9 start-page: e58039 year: 2020 ident: 98_CR35 publication-title: Elife doi: 10.7554/eLife.58039 – volume: 39 start-page: 200 year: 2018 ident: 98_CR17 publication-title: Trends Pharmacol. Sci. doi: 10.1016/j.tips.2017.11.009 – volume: 163 start-page: 1314 year: 2015 ident: 98_CR1 publication-title: Cell doi: 10.1016/j.cell.2015.11.007 – volume: 12 start-page: 174 year: 2000 ident: 98_CR31 publication-title: Curr. Opin. Cell Biol. doi: 10.1016/S0955-0674(99)00073-3 – volume: 10 start-page: 5146 year: 2020 ident: 98_CR11 publication-title: Sci. Rep. doi: 10.1038/s41598-020-61478-1 – volume: 6 start-page: e19170 year: 2011 ident: 98_CR24 publication-title: PLoS ONE doi: 10.1371/journal.pone.0019170 – volume: 28 start-page: 593 year: 2014 ident: 98_CR20 publication-title: Fundam. Clin. Pharmacol. doi: 10.1111/fcp.12080 – volume: 34 start-page: 631 year: 2016 ident: 98_CR30 publication-title: Nat. Biotechnol. doi: 10.1038/nbt.3536 – volume: 8 start-page: S7 year: 2007 ident: 98_CR27 publication-title: Genome Biol. doi: 10.1186/gb-2007-8-s1-s7 – volume: 18 start-page: 770 year: 2019 ident: 98_CR46 publication-title: Nat. Rev. Drug Discov. doi: 10.1038/s41573-019-0033-4 – volume: 7 start-page: e47826 year: 2012 ident: 98_CR42 publication-title: PLoS ONE doi: 10.1371/journal.pone.0047826 – volume: 39 start-page: 209 year: 2018 ident: 98_CR15 publication-title: Trends Pharmacol. Sci. doi: 10.1016/j.tips.2017.11.006 – volume: 76 start-page: 481 year: 2007 ident: 98_CR19 publication-title: Annu. Rev. Biochem. doi: 10.1146/annurev.biochem.76.060305.150444 – volume: 282 start-page: 34235 year: 2007 ident: 98_CR23 publication-title: J. Biol. Chem. doi: 10.1074/jbc.M706765200 – volume: 116 start-page: 10842 year: 2019 ident: 98_CR3 publication-title: Proc. Natl. Acad. Sci. doi: 10.1073/pnas.1903808116 – ident: 98_CR29 – volume: 277 start-page: 31249 year: 2002 ident: 98_CR39 publication-title: J. Biol. Chem. doi: 10.1074/jbc.M202753200 – volume: 182 start-page: 1531 year: 2020 ident: 98_CR18 publication-title: Cell doi: 10.1016/j.cell.2020.07.043 – volume: 8 start-page: 15031 year: 2017 ident: 98_CR16 publication-title: Nat. Commun. doi: 10.1038/ncomms15031 – volume: 182 start-page: 1519 year: 2020 ident: 98_CR49 publication-title: Cell doi: 10.1016/j.cell.2020.07.035 – volume: 7 start-page: 29 year: 2007 ident: 98_CR36 publication-title: Pharmacogenom. J. doi: 10.1038/sj.tpj.6500393 – volume: 95 start-page: 2976 year: 2008 ident: 98_CR8 publication-title: Biophys. J. doi: 10.1529/biophysj.108.131276 – volume: 13 start-page: 501 year: 2016 ident: 98_CR12 publication-title: Nat. Methods doi: 10.1038/nmeth.3836 – volume: 66 start-page: 241 year: 2001 ident: 98_CR32 publication-title: Prog. Nucleic Acid Res. Mol. Biol. doi: 10.1016/S0079-6603(00)66031-2 – volume: 47 start-page: 647 year: 2015 ident: 98_CR44 publication-title: Nat. Genet. doi: 10.1038/ng.3302 – volume: 66 start-page: 598 year: 2014 ident: 98_CR37 publication-title: Pharmacol. Rev. doi: 10.1124/pr.113.008219 – volume: 272 start-page: 6823 year: 1997 ident: 98_CR40 publication-title: J. Biol. Chem. doi: 10.1074/jbc.272.11.6823 – volume: 88 start-page: 645 year: 2013 ident: 98_CR13 publication-title: Biol. Rev. Camb. Philos. Soc. doi: 10.1111/brv.12020 – volume: 13 start-page: 290 year: 2014 ident: 98_CR22 publication-title: Nat. Rev. Drug Discov. doi: 10.1038/nrd4228 – volume: 115 start-page: 130 year: 2019 ident: 98_CR43 publication-title: Cardiovasc. Res. doi: 10.1093/cvr/cvy161 – volume-title: FRET and FLIM Techniques year: 2011 ident: 98_CR6 – volume: 179 start-page: 787 year: 2019 ident: 98_CR4 publication-title: Cell doi: 10.1016/j.cell.2019.09.016 – ident: 98_CR21 – volume: 14 start-page: 237 year: 2017 ident: 98_CR2 publication-title: Nat. Methods doi: 10.1038/nmeth.4196 – volume: 175 start-page: 441 year: 2006 ident: 98_CR48 publication-title: J. Cell Biol. doi: 10.1083/jcb.200605050 – volume: 11 start-page: 725 year: 2011 ident: 98_CR45 publication-title: Curr. Opin. Pharmacol. doi: 10.1016/j.coph.2011.10.006 – volume: 46 start-page: 135 year: 2018 ident: 98_CR14 publication-title: Cell. Signal. doi: 10.1016/j.cellsig.2018.03.002 – volume: 30 start-page: 304 year: 2010 ident: 98_CR34 publication-title: J. Recept. Signal Transduct. Res. doi: 10.3109/10799893.2010.509728 – volume: 470 start-page: 693 year: 2018 ident: 98_CR41 publication-title: Pflüg. Arch. Eur. J. Physiol. doi: 10.1007/s00424-017-2106-8 – volume: 11 start-page: 707 year: 2011 ident: 98_CR47 publication-title: Curr. Opin. Pharmacol. doi: 10.1016/j.coph.2011.09.001 – volume: 18 start-page: 100 year: 2021 ident: 98_CR26 publication-title: Nat. Methods doi: 10.1038/s41592-020-01018-x
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Snippet	Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to record the... Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein-protein interactions and is frequently used to record the... Abstract Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative method to screen for protein–protein interactions and is frequently used to...
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Title	Dynamic FRET-FLIM based screening of signal transduction pathways
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