--- _id: '9540' abstract: - lang: eng text: The hexameric AAA-ATPase Drg1 is a key factor in eukaryotic ribosome biogenesis and initiates cytoplasmic maturation of the large ribosomal subunit by releasing the shuttling maturation factor Rlp24. Drg1 monomers contain two AAA-domains (D1 and D2) that act in a concerted manner. Rlp24 release is inhibited by the drug diazaborine which blocks ATP hydrolysis in D2. The mode of inhibition was unknown. Here we show the first cryo-EM structure of Drg1 revealing the inhibitory mechanism. Diazaborine forms a covalent bond to the 2′-OH of the nucleotide in D2, explaining its specificity for this site. As a consequence, the D2 domain is locked in a rigid, inactive state, stalling the whole Drg1 hexamer. Resistance mechanisms identified include abolished drug binding and altered positioning of the nucleotide. Our results suggest nucleotide-modifying compounds as potential novel inhibitors for AAA-ATPases. acknowledged_ssus: - _id: EM-Fac acknowledgement: We are deeply grateful to the late Gregor Högenauer who built the foundation for this study with his visionary work on the inhibitor diazaborine and its bacterial target. We thank Rolf Breinbauer for insightful discussions on boron chemistry. We thank Anton Meinhart and Tim Clausen for the valuable discussion of the manuscript. We are indebted to Thomas Köcher for the MS measurement of the diazaborine-ATPγS adduct. We thank the team of the VBCF for support during early phases of this work and the IST Austria Electron Microscopy Facility for providing equipment. The lab of D.H. is supported by Boehringer Ingelheim. The work was funded by FWF projects P32536 and P32977 (to H.B.). article_number: '3483' article_processing_charge: No article_type: original author: - first_name: Michael full_name: Prattes, Michael last_name: Prattes - first_name: Irina full_name: Grishkovskaya, Irina last_name: Grishkovskaya - first_name: Victor-Valentin full_name: Hodirnau, Victor-Valentin id: 3661B498-F248-11E8-B48F-1D18A9856A87 last_name: Hodirnau - first_name: Ingrid full_name: Rössler, Ingrid last_name: Rössler - first_name: Isabella full_name: Klein, Isabella last_name: Klein - first_name: Christina full_name: Hetzmannseder, Christina last_name: Hetzmannseder - first_name: Gertrude full_name: Zisser, Gertrude last_name: Zisser - first_name: Christian C. full_name: Gruber, Christian C. last_name: Gruber - first_name: Karl full_name: Gruber, Karl last_name: Gruber - first_name: David full_name: Haselbach, David last_name: Haselbach - first_name: Helmut full_name: Bergler, Helmut last_name: Bergler citation: ama: Prattes M, Grishkovskaya I, Hodirnau V-V, et al. Structural basis for inhibition of the AAA-ATPase Drg1 by diazaborine. Nature Communications. 2021;12(1). doi:10.1038/s41467-021-23854-x apa: Prattes, M., Grishkovskaya, I., Hodirnau, V.-V., Rössler, I., Klein, I., Hetzmannseder, C., … Bergler, H. (2021). Structural basis for inhibition of the AAA-ATPase Drg1 by diazaborine. Nature Communications. Springer Nature. https://doi.org/10.1038/s41467-021-23854-x chicago: Prattes, Michael, Irina Grishkovskaya, Victor-Valentin Hodirnau, Ingrid Rössler, Isabella Klein, Christina Hetzmannseder, Gertrude Zisser, et al. “Structural Basis for Inhibition of the AAA-ATPase Drg1 by Diazaborine.” Nature Communications. Springer Nature, 2021. https://doi.org/10.1038/s41467-021-23854-x. ieee: M. Prattes et al., “Structural basis for inhibition of the AAA-ATPase Drg1 by diazaborine,” Nature Communications, vol. 12, no. 1. Springer Nature, 2021. ista: Prattes M, Grishkovskaya I, Hodirnau V-V, Rössler I, Klein I, Hetzmannseder C, Zisser G, Gruber CC, Gruber K, Haselbach D, Bergler H. 2021. Structural basis for inhibition of the AAA-ATPase Drg1 by diazaborine. Nature Communications. 12(1), 3483. mla: Prattes, Michael, et al. “Structural Basis for Inhibition of the AAA-ATPase Drg1 by Diazaborine.” Nature Communications, vol. 12, no. 1, 3483, Springer Nature, 2021, doi:10.1038/s41467-021-23854-x. short: M. Prattes, I. Grishkovskaya, V.-V. Hodirnau, I. Rössler, I. Klein, C. Hetzmannseder, G. Zisser, C.C. Gruber, K. Gruber, D. Haselbach, H. Bergler, Nature Communications 12 (2021). date_created: 2021-06-10T14:57:45Z date_published: 2021-06-09T00:00:00Z date_updated: 2023-08-08T14:05:26Z day: '09' ddc: - '570' department: - _id: EM-Fac doi: 10.1038/s41467-021-23854-x external_id: isi: - '000664874700014' pmid: - '34108481' file: - access_level: open_access checksum: 40fc24c1310930990b52a8ad1142ee97 content_type: application/pdf creator: cziletti date_created: 2021-06-15T18:55:59Z date_updated: 2021-06-15T18:55:59Z file_id: '9556' file_name: 2021_NatureComm_Prattes.pdf file_size: 3397292 relation: main_file success: 1 file_date_updated: 2021-06-15T18:55:59Z has_accepted_license: '1' intvolume: ' 12' isi: 1 issue: '1' keyword: - General Biochemistry - Genetics and Molecular Biology - General Physics and Astronomy - General Chemistry language: - iso: eng month: '06' oa: 1 oa_version: Published Version pmid: 1 publication: Nature Communications publication_identifier: eissn: - 2041-1723 publication_status: published publisher: Springer Nature quality_controlled: '1' status: public title: Structural basis for inhibition of the AAA-ATPase Drg1 by diazaborine tmp: image: /images/cc_by.png legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0) short: CC BY (4.0) type: journal_article user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8 volume: 12 year: '2021' ... --- _id: '9607' abstract: - lang: eng text: While high risk of failure is an inherent part of developing innovative therapies, it can be reduced by adherence to evidence-based rigorous research practices. Numerous analyses conducted to date have clearly identified measures that need to be taken to improve research rigor. Supported through the European Union's Innovative Medicines Initiative, the EQIPD consortium has developed a novel preclinical research quality system that can be applied in both public and private sectors and is free for anyone to use. The EQIPD Quality System was designed to be suited to boost innovation by ensuring the generation of robust and reliable preclinical data while being lean, effective and not becoming a burden that could negatively impact the freedom to explore scientific questions. EQIPD defines research quality as the extent to which research data are fit for their intended use. Fitness, in this context, is defined by the stakeholders, who are the scientists directly involved in the research, but also their funders, sponsors, publishers, research tool manufacturers and collaboration partners such as peers in a multi-site research project. The essence of the EQIPD Quality System is the set of 18 core requirements that can be addressed flexibly, according to user-specific needs and following a user-defined trajectory. The EQIPD Quality System proposes guidance on expectations for quality-related measures, defines criteria for adequate processes (i.e., performance standards) and provides examples of how such measures can be developed and implemented. However, it does not prescribe any pre-determined solutions. EQIPD has also developed tools (for optional use) to support users in implementing the system and assessment services for those research units that successfully implement the quality system and seek formal accreditation. Building upon the feedback from users and continuous improvement, a sustainable EQIPD Quality System will ultimately serve the entire community of scientists conducting non-regulated preclinical research, by helping them generate reliable data that are fit for their intended use. acknowledgement: This project has received funding from the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement No 777364. This Joint Undertaking receives support from the European Union’s Horizon 2020 research and innovation programme and EFPIA. The authors are very grateful to Martin Heinrich (Abbvie, Ludwigshafen, Germany) for the exceptional IT support and programming the EQIPD Planning Tool and the Creator Tool and to Dr Shai Silberberg (NINDS, USA), Dr. Renza Roncarati (PAASP Italy) and Dr Judith Homberg (Radboud University, Nijmegen) for highly stimulating contributions to the discussions and comments on earlier versions of this manuscript. We also wish to express our thanks to Dr. Sara Stöber (concentris research management GmbH, Fürstenfeldbruck, Germany) for excellent and continuous support of this project. Creation of the EQIPD Stakeholder group was supported by Noldus Information Technology bv (Wageningen, the Netherlands). article_processing_charge: No article_type: original author: - first_name: Anton full_name: Bespalov, Anton last_name: Bespalov - first_name: René full_name: Bernard, René last_name: Bernard - first_name: Anja full_name: Gilis, Anja last_name: Gilis - first_name: Björn full_name: Gerlach, Björn last_name: Gerlach - first_name: Javier full_name: Guillén, Javier last_name: Guillén - first_name: Vincent full_name: Castagné, Vincent last_name: Castagné - first_name: Isabel A. full_name: Lefevre, Isabel A. last_name: Lefevre - first_name: Fiona full_name: Ducrey, Fiona last_name: Ducrey - first_name: Lee full_name: Monk, Lee last_name: Monk - first_name: Sandrine full_name: Bongiovanni, Sandrine last_name: Bongiovanni - first_name: Bruce full_name: Altevogt, Bruce last_name: Altevogt - first_name: María full_name: Arroyo-Araujo, María last_name: Arroyo-Araujo - first_name: Lior full_name: Bikovski, Lior last_name: Bikovski - first_name: Natasja full_name: De Bruin, Natasja last_name: De Bruin - first_name: Esmeralda full_name: Castaños-Vélez, Esmeralda last_name: Castaños-Vélez - first_name: Alexander full_name: Dityatev, Alexander last_name: Dityatev - first_name: Christoph H. full_name: Emmerich, Christoph H. last_name: Emmerich - first_name: Raafat full_name: Fares, Raafat last_name: Fares - first_name: Chantelle full_name: Ferland-Beckham, Chantelle last_name: Ferland-Beckham - first_name: Christelle full_name: Froger-Colléaux, Christelle last_name: Froger-Colléaux - first_name: Valerie full_name: Gailus-Durner, Valerie last_name: Gailus-Durner - first_name: Sabine M. full_name: Hölter, Sabine M. last_name: Hölter - first_name: Martine Cj full_name: Hofmann, Martine Cj last_name: Hofmann - first_name: Patricia full_name: Kabitzke, Patricia last_name: Kabitzke - first_name: Martien Jh full_name: Kas, Martien Jh last_name: Kas - first_name: Claudia full_name: Kurreck, Claudia last_name: Kurreck - first_name: Paul full_name: Moser, Paul last_name: Moser - first_name: Malgorzata full_name: Pietraszek, Malgorzata last_name: Pietraszek - first_name: Piotr full_name: Popik, Piotr last_name: Popik - first_name: Heidrun full_name: Potschka, Heidrun last_name: Potschka - first_name: Ernesto full_name: Prado Montes De Oca, Ernesto last_name: Prado Montes De Oca - first_name: Leonardo full_name: Restivo, Leonardo last_name: Restivo - first_name: Gernot full_name: Riedel, Gernot last_name: Riedel - first_name: Merel full_name: Ritskes-Hoitinga, Merel last_name: Ritskes-Hoitinga - first_name: Janko full_name: Samardzic, Janko last_name: Samardzic - first_name: Michael full_name: Schunn, Michael id: 4272DB4A-F248-11E8-B48F-1D18A9856A87 last_name: Schunn orcid: 0000-0003-4326-5300 - first_name: Claudia full_name: Stöger, Claudia last_name: Stöger - first_name: Vootele full_name: Voikar, Vootele last_name: Voikar - first_name: Jan full_name: Vollert, Jan last_name: Vollert - first_name: Kimberley E. full_name: Wever, Kimberley E. last_name: Wever - first_name: Kathleen full_name: Wuyts, Kathleen last_name: Wuyts - first_name: Malcolm R. full_name: Macleod, Malcolm R. last_name: Macleod - first_name: Ulrich full_name: Dirnagl, Ulrich last_name: Dirnagl - first_name: Thomas full_name: Steckler, Thomas last_name: Steckler citation: ama: Bespalov A, Bernard R, Gilis A, et al. Introduction to the EQIPD quality system. eLife. 2021;10. doi:10.7554/eLife.63294 apa: Bespalov, A., Bernard, R., Gilis, A., Gerlach, B., Guillén, J., Castagné, V., … Steckler, T. (2021). Introduction to the EQIPD quality system. ELife. eLife Sciences Publications. https://doi.org/10.7554/eLife.63294 chicago: Bespalov, Anton, René Bernard, Anja Gilis, Björn Gerlach, Javier Guillén, Vincent Castagné, Isabel A. Lefevre, et al. “Introduction to the EQIPD Quality System.” ELife. eLife Sciences Publications, 2021. https://doi.org/10.7554/eLife.63294. ieee: A. Bespalov et al., “Introduction to the EQIPD quality system,” eLife, vol. 10. eLife Sciences Publications, 2021. ista: Bespalov A, Bernard R, Gilis A, Gerlach B, Guillén J, Castagné V, Lefevre IA, Ducrey F, Monk L, Bongiovanni S, Altevogt B, Arroyo-Araujo M, Bikovski L, De Bruin N, Castaños-Vélez E, Dityatev A, Emmerich CH, Fares R, Ferland-Beckham C, Froger-Colléaux C, Gailus-Durner V, Hölter SM, Hofmann MC, Kabitzke P, Kas MJ, Kurreck C, Moser P, Pietraszek M, Popik P, Potschka H, Prado Montes De Oca E, Restivo L, Riedel G, Ritskes-Hoitinga M, Samardzic J, Schunn M, Stöger C, Voikar V, Vollert J, Wever KE, Wuyts K, Macleod MR, Dirnagl U, Steckler T. 2021. Introduction to the EQIPD quality system. eLife. 10. mla: Bespalov, Anton, et al. “Introduction to the EQIPD Quality System.” ELife, vol. 10, eLife Sciences Publications, 2021, doi:10.7554/eLife.63294. short: A. Bespalov, R. Bernard, A. Gilis, B. Gerlach, J. Guillén, V. Castagné, I.A. Lefevre, F. Ducrey, L. Monk, S. Bongiovanni, B. Altevogt, M. Arroyo-Araujo, L. Bikovski, N. De Bruin, E. Castaños-Vélez, A. Dityatev, C.H. Emmerich, R. Fares, C. Ferland-Beckham, C. Froger-Colléaux, V. Gailus-Durner, S.M. Hölter, M.C. Hofmann, P. Kabitzke, M.J. Kas, C. Kurreck, P. Moser, M. Pietraszek, P. Popik, H. Potschka, E. Prado Montes De Oca, L. Restivo, G. Riedel, M. Ritskes-Hoitinga, J. Samardzic, M. Schunn, C. Stöger, V. Voikar, J. Vollert, K.E. Wever, K. Wuyts, M.R. Macleod, U. Dirnagl, T. Steckler, ELife 10 (2021). date_created: 2021-06-27T22:01:49Z date_published: 2021-05-24T00:00:00Z date_updated: 2023-08-10T13:36:50Z day: '24' ddc: - '570' department: - _id: PreCl doi: 10.7554/eLife.63294 external_id: isi: - '000661272000001' pmid: - '34028353' file: - access_level: open_access checksum: 885b746051a7a6b6e24e3d2781a48fde content_type: application/pdf creator: asandaue date_created: 2021-06-28T11:35:30Z date_updated: 2021-06-28T11:35:30Z file_id: '9609' file_name: 2021_ELife_Bespalov.pdf file_size: 2500720 relation: main_file success: 1 file_date_updated: 2021-06-28T11:35:30Z has_accepted_license: '1' intvolume: ' 10' isi: 1 language: - iso: eng month: '05' oa: 1 oa_version: Published Version pmid: 1 publication: eLife publication_identifier: eissn: - 2050084X publication_status: published publisher: eLife Sciences Publications quality_controlled: '1' scopus_import: '1' status: public title: Introduction to the EQIPD quality system tmp: image: /images/cc_by.png legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0) short: CC BY (4.0) type: journal_article user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8 volume: 10 year: '2021' ... --- _id: '9603' abstract: - lang: eng text: Mosaic analysis with double markers (MADM) offers one approach to visualize and concomitantly manipulate genetically defined cells in mice with single-cell resolution. MADM applications include the analysis of lineage, single-cell morphology and physiology, genomic imprinting phenotypes, and dissection of cell-autonomous gene functions in vivo in health and disease. Yet, MADM can only be applied to <25% of all mouse genes on select chromosomes to date. To overcome this limitation, we generate transgenic mice with knocked-in MADM cassettes near the centromeres of all 19 autosomes and validate their use across organs. With this resource, >96% of the entire mouse genome can now be subjected to single-cell genetic mosaic analysis. Beyond a proof of principle, we apply our MADM library to systematically trace sister chromatid segregation in distinct mitotic cell lineages. We find striking chromosome-specific biases in segregation patterns, reflecting a putative mechanism for the asymmetric segregation of genetic determinants in somatic stem cell division. acknowledged_ssus: - _id: Bio - _id: LifeSc - _id: PreCl acknowledgement: We thank the Bioimaging, Life Science, and Pre-Clinical Facilities at IST Austria; M.P. Postiglione, C. Simbriger, K. Valoskova, C. Schwayer, T. Hussain, M. Pieber, and V. Wimmer for initial experiments, technical support, and/or assistance; R. Shigemoto for sharing iv (Dnah11 mutant) mice; and M. Sixt and all members of the Hippenmeyer lab for discussion. This work was supported by National Institutes of Health grants ( R01-NS050580 to L.L. and F32MH096361 to L.A.S.). L.L. is an investigator of HHMI. N.A. received support from FWF Firnberg-Programm ( T 1031 ). A.H.H. is a recipient of a DOC Fellowship (24812) of the Austrian Academy of Sciences . This work also received support from IST Austria institutional funds , FWF SFB F78 to S.H., the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme ( FP7/2007-2013 ) under REA grant agreement no 618444 to S.H., and the European Research Council (ERC) under the European Union’s Horizon 2020 Research and Innovation Programme (grant agreement no. 725780 LinPro ) to S.H. article_number: '109274' article_processing_charge: No article_type: original author: - first_name: Ximena full_name: Contreras, Ximena id: 475990FE-F248-11E8-B48F-1D18A9856A87 last_name: Contreras - first_name: Nicole full_name: Amberg, Nicole id: 4CD6AAC6-F248-11E8-B48F-1D18A9856A87 last_name: Amberg orcid: 0000-0002-3183-8207 - first_name: Amarbayasgalan full_name: Davaatseren, Amarbayasgalan id: 70ADC922-B424-11E9-99E3-BA18E6697425 last_name: Davaatseren - first_name: Andi H full_name: Hansen, Andi H id: 38853E16-F248-11E8-B48F-1D18A9856A87 last_name: Hansen - first_name: Johanna full_name: Sonntag, Johanna id: 32FE7D7C-F248-11E8-B48F-1D18A9856A87 last_name: Sonntag - first_name: Lill full_name: Andersen, Lill last_name: Andersen - first_name: Tina full_name: Bernthaler, Tina last_name: Bernthaler - first_name: Carmen full_name: Streicher, Carmen id: 36BCB99C-F248-11E8-B48F-1D18A9856A87 last_name: Streicher - first_name: Anna-Magdalena full_name: Heger, Anna-Magdalena id: 4B76FFD2-F248-11E8-B48F-1D18A9856A87 last_name: Heger - first_name: Randy L. full_name: Johnson, Randy L. last_name: Johnson - first_name: Lindsay A. full_name: Schwarz, Lindsay A. last_name: Schwarz - first_name: Liqun full_name: Luo, Liqun last_name: Luo - first_name: Thomas full_name: Rülicke, Thomas last_name: Rülicke - first_name: Simon full_name: Hippenmeyer, Simon id: 37B36620-F248-11E8-B48F-1D18A9856A87 last_name: Hippenmeyer orcid: 0000-0003-2279-1061 citation: ama: Contreras X, Amberg N, Davaatseren A, et al. A genome-wide library of MADM mice for single-cell genetic mosaic analysis. Cell Reports. 2021;35(12). doi:10.1016/j.celrep.2021.109274 apa: Contreras, X., Amberg, N., Davaatseren, A., Hansen, A. H., Sonntag, J., Andersen, L., … Hippenmeyer, S. (2021). A genome-wide library of MADM mice for single-cell genetic mosaic analysis. Cell Reports. Cell Press. https://doi.org/10.1016/j.celrep.2021.109274 chicago: Contreras, Ximena, Nicole Amberg, Amarbayasgalan Davaatseren, Andi H Hansen, Johanna Sonntag, Lill Andersen, Tina Bernthaler, et al. “A Genome-Wide Library of MADM Mice for Single-Cell Genetic Mosaic Analysis.” Cell Reports. Cell Press, 2021. https://doi.org/10.1016/j.celrep.2021.109274. ieee: X. Contreras et al., “A genome-wide library of MADM mice for single-cell genetic mosaic analysis,” Cell Reports, vol. 35, no. 12. Cell Press, 2021. ista: Contreras X, Amberg N, Davaatseren A, Hansen AH, Sonntag J, Andersen L, Bernthaler T, Streicher C, Heger A-M, Johnson RL, Schwarz LA, Luo L, Rülicke T, Hippenmeyer S. 2021. A genome-wide library of MADM mice for single-cell genetic mosaic analysis. Cell Reports. 35(12), 109274. mla: Contreras, Ximena, et al. “A Genome-Wide Library of MADM Mice for Single-Cell Genetic Mosaic Analysis.” Cell Reports, vol. 35, no. 12, 109274, Cell Press, 2021, doi:10.1016/j.celrep.2021.109274. short: X. Contreras, N. Amberg, A. Davaatseren, A.H. Hansen, J. Sonntag, L. Andersen, T. Bernthaler, C. Streicher, A.-M. Heger, R.L. Johnson, L.A. Schwarz, L. Luo, T. Rülicke, S. Hippenmeyer, Cell Reports 35 (2021). date_created: 2021-06-27T22:01:48Z date_published: 2021-06-22T00:00:00Z date_updated: 2023-08-10T13:55:00Z day: '22' ddc: - '570' department: - _id: SiHi - _id: LoSw - _id: PreCl doi: 10.1016/j.celrep.2021.109274 ec_funded: 1 external_id: isi: - '000664463600016' file: - access_level: open_access checksum: d49520fdcbbb5c2f883bddb67cee5d77 content_type: application/pdf creator: asandaue date_created: 2021-06-28T14:06:24Z date_updated: 2021-06-28T14:06:24Z file_id: '9613' file_name: 2021_CellReports_Contreras.pdf file_size: 7653149 relation: main_file success: 1 file_date_updated: 2021-06-28T14:06:24Z has_accepted_license: '1' intvolume: ' 35' isi: 1 issue: '12' language: - iso: eng month: '06' oa: 1 oa_version: Published Version project: - _id: 2625A13E-B435-11E9-9278-68D0E5697425 grant_number: '24812' name: Molecular Mechanisms of Radial Neuronal Migration - _id: 25D61E48-B435-11E9-9278-68D0E5697425 call_identifier: FP7 grant_number: '618444' name: Molecular Mechanisms of Cerebral Cortex Development - _id: 260018B0-B435-11E9-9278-68D0E5697425 call_identifier: H2020 grant_number: '725780' name: Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development publication: Cell Reports publication_identifier: eissn: - '22111247' publication_status: published publisher: Cell Press quality_controlled: '1' related_material: link: - description: News on IST Homepage relation: press_release url: https://ist.ac.at/en/news/boost-for-mouse-genetic-analysis/ scopus_import: '1' status: public title: A genome-wide library of MADM mice for single-cell genetic mosaic analysis tmp: image: /images/cc_by_nc_nd.png legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) short: CC BY-NC-ND (4.0) type: journal_article user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8 volume: 35 year: '2021' ... --- _id: '9822' abstract: - lang: eng text: Attachment of adhesive molecules on cell culture surfaces to restrict cell adhesion to defined areas and shapes has been vital for the progress of in vitro research. In currently existing patterning methods, a combination of pattern properties such as stability, precision, specificity, high-throughput outcome, and spatiotemporal control is highly desirable but challenging to achieve. Here, we introduce a versatile and high-throughput covalent photoimmobilization technique, comprising a light-dose-dependent patterning step and a subsequent functionalization of the pattern via click chemistry. This two-step process is feasible on arbitrary surfaces and allows for generation of sustainable patterns and gradients. The method is validated in different biological systems by patterning adhesive ligands on cell-repellent surfaces, thereby constraining the growth and migration of cells to the designated areas. We then implement a sequential photopatterning approach by adding a second switchable patterning step, allowing for spatiotemporal control over two distinct surface patterns. As a proof of concept, we reconstruct the dynamics of the tip/stalk cell switch during angiogenesis. Our results show that the spatiotemporal control provided by our “sequential photopatterning” system is essential for mimicking dynamic biological processes and that our innovative approach has great potential for further applications in cell science. acknowledgement: We would like to thank Charlott Leu for the production of our chromium wafers, Louise Ritter for her contribution of the IF stainings in Figure 4, Shokoufeh Teymouri for her help with the Bioinert coated slides, and finally Prof. Dr. Joachim Rädler for his valuable scientific guidance. article_processing_charge: Yes (in subscription journal) article_type: original author: - first_name: Themistoklis full_name: Zisis, Themistoklis last_name: Zisis - first_name: Jan full_name: Schwarz, Jan id: 346C1EC6-F248-11E8-B48F-1D18A9856A87 last_name: Schwarz - first_name: Miriam full_name: Balles, Miriam last_name: Balles - first_name: Maibritt full_name: Kretschmer, Maibritt last_name: Kretschmer - first_name: Maria full_name: Nemethova, Maria id: 34E27F1C-F248-11E8-B48F-1D18A9856A87 last_name: Nemethova - first_name: Remy P full_name: Chait, Remy P id: 3464AE84-F248-11E8-B48F-1D18A9856A87 last_name: Chait orcid: 0000-0003-0876-3187 - first_name: Robert full_name: Hauschild, Robert id: 4E01D6B4-F248-11E8-B48F-1D18A9856A87 last_name: Hauschild orcid: 0000-0001-9843-3522 - first_name: Janina full_name: Lange, Janina last_name: Lange - first_name: Calin C full_name: Guet, Calin C id: 47F8433E-F248-11E8-B48F-1D18A9856A87 last_name: Guet orcid: 0000-0001-6220-2052 - first_name: Michael K full_name: Sixt, Michael K id: 41E9FBEA-F248-11E8-B48F-1D18A9856A87 last_name: Sixt orcid: 0000-0002-4561-241X - first_name: Stefan full_name: Zahler, Stefan last_name: Zahler citation: ama: Zisis T, Schwarz J, Balles M, et al. Sequential and switchable patterning for studying cellular processes under spatiotemporal control. ACS Applied Materials and Interfaces. 2021;13(30):35545–35560. doi:10.1021/acsami.1c09850 apa: Zisis, T., Schwarz, J., Balles, M., Kretschmer, M., Nemethova, M., Chait, R. P., … Zahler, S. (2021). Sequential and switchable patterning for studying cellular processes under spatiotemporal control. ACS Applied Materials and Interfaces. American Chemical Society. https://doi.org/10.1021/acsami.1c09850 chicago: Zisis, Themistoklis, Jan Schwarz, Miriam Balles, Maibritt Kretschmer, Maria Nemethova, Remy P Chait, Robert Hauschild, et al. “Sequential and Switchable Patterning for Studying Cellular Processes under Spatiotemporal Control.” ACS Applied Materials and Interfaces. American Chemical Society, 2021. https://doi.org/10.1021/acsami.1c09850. ieee: T. Zisis et al., “Sequential and switchable patterning for studying cellular processes under spatiotemporal control,” ACS Applied Materials and Interfaces, vol. 13, no. 30. American Chemical Society, pp. 35545–35560, 2021. ista: Zisis T, Schwarz J, Balles M, Kretschmer M, Nemethova M, Chait RP, Hauschild R, Lange J, Guet CC, Sixt MK, Zahler S. 2021. Sequential and switchable patterning for studying cellular processes under spatiotemporal control. ACS Applied Materials and Interfaces. 13(30), 35545–35560. mla: Zisis, Themistoklis, et al. “Sequential and Switchable Patterning for Studying Cellular Processes under Spatiotemporal Control.” ACS Applied Materials and Interfaces, vol. 13, no. 30, American Chemical Society, 2021, pp. 35545–35560, doi:10.1021/acsami.1c09850. short: T. Zisis, J. Schwarz, M. Balles, M. Kretschmer, M. Nemethova, R.P. Chait, R. Hauschild, J. Lange, C.C. Guet, M.K. Sixt, S. Zahler, ACS Applied Materials and Interfaces 13 (2021) 35545–35560. date_created: 2021-08-08T22:01:28Z date_published: 2021-08-04T00:00:00Z date_updated: 2023-08-10T14:22:48Z day: '04' ddc: - '620' - '570' department: - _id: MiSi - _id: GaTk - _id: Bio - _id: CaGu doi: 10.1021/acsami.1c09850 ec_funded: 1 external_id: isi: - '000683741400026' pmid: - '34283577' file: - access_level: open_access checksum: b043a91d9f9200e467b970b692687ed3 content_type: application/pdf creator: asandaue date_created: 2021-08-09T09:44:03Z date_updated: 2021-08-09T09:44:03Z file_id: '9833' file_name: 2021_ACSAppliedMaterialsAndInterfaces_Zisis.pdf file_size: 7123293 relation: main_file success: 1 file_date_updated: 2021-08-09T09:44:03Z has_accepted_license: '1' intvolume: ' 13' isi: 1 issue: '30' language: - iso: eng month: '08' oa: 1 oa_version: Published Version page: 35545–35560 pmid: 1 project: - _id: 25FE9508-B435-11E9-9278-68D0E5697425 call_identifier: H2020 grant_number: '724373' name: Cellular navigation along spatial gradients publication: ACS Applied Materials and Interfaces publication_identifier: eissn: - '19448252' issn: - '19448244' publication_status: published publisher: American Chemical Society quality_controlled: '1' scopus_import: '1' status: public title: Sequential and switchable patterning for studying cellular processes under spatiotemporal control tmp: image: /images/cc_by_nc_nd.png legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) short: CC BY-NC-ND (4.0) type: journal_article user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8 volume: 13 year: '2021' ... --- _id: '9911' abstract: - lang: eng text: A modern day light microscope has evolved from a tool devoted to making primarily empirical observations to what is now a sophisticated , quantitative device that is an integral part of both physical and life science research. Nowadays, microscopes are found in nearly every experimental laboratory. However, despite their prevalent use in capturing and quantifying scientific phenomena, neither a thorough understanding of the principles underlying quantitative imaging techniques nor appropriate knowledge of how to calibrate, operate and maintain microscopes can be taken for granted. This is clearly demonstrated by the well-documented and widespread difficulties that are routinely encountered in evaluating acquired data and reproducing scientific experiments. Indeed, studies have shown that more than 70% of researchers have tried and failed to repeat another scientist's experiments, while more than half have even failed to reproduce their own experiments. One factor behind the reproducibility crisis of experiments published in scientific journals is the frequent underreporting of imaging methods caused by a lack of awareness and/or a lack of knowledge of the applied technique. Whereas quality control procedures for some methods used in biomedical research, such as genomics (e.g. DNA sequencing, RNA-seq) or cytometry, have been introduced (e.g. ENCODE), this issue has not been tackled for optical microscopy instrumentation and images. Although many calibration standards and protocols have been published, there is a lack of awareness and agreement on common standards and guidelines for quality assessment and reproducibility. In April 2020, the QUality Assessment and REProducibility for instruments and images in Light Microscopy (QUAREP-LiMi) initiative was formed. This initiative comprises imaging scientists from academia and industry who share a common interest in achieving a better understanding of the performance and limitations of microscopes and improved quality control (QC) in light microscopy. The ultimate goal of the QUAREP-LiMi initiative is to establish a set of common QC standards, guidelines, metadata models and tools, including detailed protocols, with the ultimate aim of improving reproducible advances in scientific research. This White Paper (1) summarizes the major obstacles identified in the field that motivated the launch of the QUAREP-LiMi initiative; (2) identifies the urgent need to address these obstacles in a grassroots manner, through a community of stakeholders including, researchers, imaging scientists, bioimage analysts, bioimage informatics developers, corporate partners, funding agencies, standards organizations, scientific publishers and observers of such; (3) outlines the current actions of the QUAREP-LiMi initiative and (4) proposes future steps that can be taken to improve the dissemination and acceptance of the proposed guidelines to manage QC. To summarize, the principal goal of the QUAREP-LiMi initiative is to improve the overall quality and reproducibility of light microscope image data by introducing broadly accepted standard practices and accurately captured image data metrics. acknowledgement: We thank https://www.somersault1824.com/somersault18:24 BV (Leuven, Belgium) for help with Figure 1. E. C.-S. was supported by the project PPBI-POCI-01-0145-FEDER-022122, in the scope of Fundação para a Ciência e Tecnologia, Portugal (FCT) National Roadmap of Research Infrastructures. R.N. was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) Grant number Ni 451/9-1 - MIAP-Freiburg. article_processing_charge: Yes article_type: original author: - first_name: Glyn full_name: Nelson, Glyn last_name: Nelson - first_name: Ulrike full_name: Boehm, Ulrike last_name: Boehm - first_name: Steve full_name: Bagley, Steve last_name: Bagley - first_name: Peter full_name: Bajcsy, Peter last_name: Bajcsy - first_name: Johanna full_name: Bischof, Johanna last_name: Bischof - first_name: Claire M. full_name: Brown, Claire M. last_name: Brown - first_name: Aurélien full_name: Dauphin, Aurélien last_name: Dauphin - first_name: Ian M. full_name: Dobbie, Ian M. last_name: Dobbie - first_name: John E. full_name: Eriksson, John E. last_name: Eriksson - first_name: Orestis full_name: Faklaris, Orestis last_name: Faklaris - first_name: Julia full_name: Fernandez-Rodriguez, Julia last_name: Fernandez-Rodriguez - first_name: Alexia full_name: Ferrand, Alexia last_name: Ferrand - first_name: Laurent full_name: Gelman, Laurent last_name: Gelman - first_name: Ali full_name: Gheisari, Ali last_name: Gheisari - first_name: Hella full_name: Hartmann, Hella last_name: Hartmann - first_name: Christian full_name: Kukat, Christian last_name: Kukat - first_name: Alex full_name: Laude, Alex last_name: Laude - first_name: Miso full_name: Mitkovski, Miso last_name: Mitkovski - first_name: Sebastian full_name: Munck, Sebastian last_name: Munck - first_name: Alison J. full_name: North, Alison J. last_name: North - first_name: Tobias M. full_name: Rasse, Tobias M. last_name: Rasse - first_name: Ute full_name: Resch-Genger, Ute last_name: Resch-Genger - first_name: Lucas C. full_name: Schuetz, Lucas C. last_name: Schuetz - first_name: Arne full_name: Seitz, Arne last_name: Seitz - first_name: Caterina full_name: Strambio-De-Castillia, Caterina last_name: Strambio-De-Castillia - first_name: Jason R. full_name: Swedlow, Jason R. last_name: Swedlow - first_name: Ioannis full_name: Alexopoulos, Ioannis last_name: Alexopoulos - first_name: Karin full_name: Aumayr, Karin last_name: Aumayr - first_name: Sergiy full_name: Avilov, Sergiy last_name: Avilov - first_name: Gert Jan full_name: Bakker, Gert Jan last_name: Bakker - first_name: Rodrigo R. full_name: Bammann, Rodrigo R. last_name: Bammann - first_name: Andrea full_name: Bassi, Andrea last_name: Bassi - first_name: Hannes full_name: Beckert, Hannes last_name: Beckert - first_name: Sebastian full_name: Beer, Sebastian last_name: Beer - first_name: Yury full_name: Belyaev, Yury last_name: Belyaev - first_name: Jakob full_name: Bierwagen, Jakob last_name: Bierwagen - first_name: Konstantin A. full_name: Birngruber, Konstantin A. last_name: Birngruber - first_name: Manel full_name: Bosch, Manel last_name: Bosch - first_name: Juergen full_name: Breitlow, Juergen last_name: Breitlow - first_name: Lisa A. full_name: Cameron, Lisa A. last_name: Cameron - first_name: Joe full_name: Chalfoun, Joe last_name: Chalfoun - first_name: James J. full_name: Chambers, James J. last_name: Chambers - first_name: Chieh Li full_name: Chen, Chieh Li last_name: Chen - first_name: Eduardo full_name: Conde-Sousa, Eduardo last_name: Conde-Sousa - first_name: Alexander D. full_name: Corbett, Alexander D. last_name: Corbett - first_name: Fabrice P. full_name: Cordelieres, Fabrice P. last_name: Cordelieres - first_name: Elaine Del full_name: Nery, Elaine Del last_name: Nery - first_name: Ralf full_name: Dietzel, Ralf last_name: Dietzel - first_name: Frank full_name: Eismann, Frank last_name: Eismann - first_name: Elnaz full_name: Fazeli, Elnaz last_name: Fazeli - first_name: Andreas full_name: Felscher, Andreas last_name: Felscher - first_name: Hans full_name: Fried, Hans last_name: Fried - first_name: Nathalie full_name: Gaudreault, Nathalie last_name: Gaudreault - first_name: Wah Ing full_name: Goh, Wah Ing last_name: Goh - first_name: Thomas full_name: Guilbert, Thomas last_name: Guilbert - first_name: Roland full_name: Hadleigh, Roland last_name: Hadleigh - first_name: Peter full_name: Hemmerich, Peter last_name: Hemmerich - first_name: Gerhard A. full_name: Holst, Gerhard A. last_name: Holst - first_name: Michelle S. full_name: Itano, Michelle S. last_name: Itano - first_name: Claudia B. full_name: Jaffe, Claudia B. last_name: Jaffe - first_name: Helena K. full_name: Jambor, Helena K. last_name: Jambor - first_name: Stuart C. full_name: Jarvis, Stuart C. last_name: Jarvis - first_name: Antje full_name: Keppler, Antje last_name: Keppler - first_name: David full_name: Kirchenbuechler, David last_name: Kirchenbuechler - first_name: Marcel full_name: Kirchner, Marcel last_name: Kirchner - first_name: Norio full_name: Kobayashi, Norio last_name: Kobayashi - first_name: Gabriel full_name: Krens, Gabriel id: 2B819732-F248-11E8-B48F-1D18A9856A87 last_name: Krens orcid: 0000-0003-4761-5996 - first_name: Susanne full_name: Kunis, Susanne last_name: Kunis - first_name: Judith full_name: Lacoste, Judith last_name: Lacoste - first_name: Marco full_name: Marcello, Marco last_name: Marcello - first_name: Gabriel G. full_name: Martins, Gabriel G. last_name: Martins - first_name: Daniel J. full_name: Metcalf, Daniel J. last_name: Metcalf - first_name: Claire A. full_name: Mitchell, Claire A. last_name: Mitchell - first_name: Joshua full_name: Moore, Joshua last_name: Moore - first_name: Tobias full_name: Mueller, Tobias last_name: Mueller - first_name: Michael S. full_name: Nelson, Michael S. last_name: Nelson - first_name: Stephen full_name: Ogg, Stephen last_name: Ogg - first_name: Shuichi full_name: Onami, Shuichi last_name: Onami - first_name: Alexandra L. full_name: Palmer, Alexandra L. last_name: Palmer - first_name: Perrine full_name: Paul-Gilloteaux, Perrine last_name: Paul-Gilloteaux - first_name: Jaime A. full_name: Pimentel, Jaime A. last_name: Pimentel - first_name: Laure full_name: Plantard, Laure last_name: Plantard - first_name: Santosh full_name: Podder, Santosh last_name: Podder - first_name: Elton full_name: Rexhepaj, Elton last_name: Rexhepaj - first_name: Arnaud full_name: Royon, Arnaud last_name: Royon - first_name: Markku A. full_name: Saari, Markku A. last_name: Saari - first_name: Damien full_name: Schapman, Damien last_name: Schapman - first_name: Vincent full_name: Schoonderwoert, Vincent last_name: Schoonderwoert - first_name: Britta full_name: Schroth-Diez, Britta last_name: Schroth-Diez - first_name: Stanley full_name: Schwartz, Stanley last_name: Schwartz - first_name: Michael full_name: Shaw, Michael last_name: Shaw - first_name: Martin full_name: Spitaler, Martin last_name: Spitaler - first_name: Martin T. full_name: Stoeckl, Martin T. last_name: Stoeckl - first_name: Damir full_name: Sudar, Damir last_name: Sudar - first_name: Jeremie full_name: Teillon, Jeremie last_name: Teillon - first_name: Stefan full_name: Terjung, Stefan last_name: Terjung - first_name: Roland full_name: Thuenauer, Roland last_name: Thuenauer - first_name: Christian D. full_name: Wilms, Christian D. last_name: Wilms - first_name: Graham D. full_name: Wright, Graham D. last_name: Wright - first_name: Roland full_name: Nitschke, Roland last_name: Nitschke citation: ama: 'Nelson G, Boehm U, Bagley S, et al. QUAREP-LiMi: A community-driven initiative to establish guidelines for quality assessment and reproducibility for instruments and images in light microscopy. Journal of Microscopy. 2021;284(1):56-73. doi:10.1111/jmi.13041' apa: 'Nelson, G., Boehm, U., Bagley, S., Bajcsy, P., Bischof, J., Brown, C. M., … Nitschke, R. (2021). QUAREP-LiMi: A community-driven initiative to establish guidelines for quality assessment and reproducibility for instruments and images in light microscopy. Journal of Microscopy. Wiley. https://doi.org/10.1111/jmi.13041' chicago: 'Nelson, Glyn, Ulrike Boehm, Steve Bagley, Peter Bajcsy, Johanna Bischof, Claire M. Brown, Aurélien Dauphin, et al. “QUAREP-LiMi: A Community-Driven Initiative to Establish Guidelines for Quality Assessment and Reproducibility for Instruments and Images in Light Microscopy.” Journal of Microscopy. Wiley, 2021. https://doi.org/10.1111/jmi.13041.' ieee: 'G. Nelson et al., “QUAREP-LiMi: A community-driven initiative to establish guidelines for quality assessment and reproducibility for instruments and images in light microscopy,” Journal of Microscopy, vol. 284, no. 1. Wiley, pp. 56–73, 2021.' ista: 'Nelson G et al. 2021. QUAREP-LiMi: A community-driven initiative to establish guidelines for quality assessment and reproducibility for instruments and images in light microscopy. Journal of Microscopy. 284(1), 56–73.' mla: 'Nelson, Glyn, et al. “QUAREP-LiMi: A Community-Driven Initiative to Establish Guidelines for Quality Assessment and Reproducibility for Instruments and Images in Light Microscopy.” Journal of Microscopy, vol. 284, no. 1, Wiley, 2021, pp. 56–73, doi:10.1111/jmi.13041.' short: G. Nelson, U. Boehm, S. Bagley, P. Bajcsy, J. Bischof, C.M. Brown, A. Dauphin, I.M. Dobbie, J.E. Eriksson, O. Faklaris, J. Fernandez-Rodriguez, A. Ferrand, L. Gelman, A. Gheisari, H. Hartmann, C. Kukat, A. Laude, M. Mitkovski, S. Munck, A.J. North, T.M. Rasse, U. Resch-Genger, L.C. Schuetz, A. Seitz, C. Strambio-De-Castillia, J.R. Swedlow, I. Alexopoulos, K. Aumayr, S. Avilov, G.J. Bakker, R.R. Bammann, A. Bassi, H. Beckert, S. Beer, Y. Belyaev, J. Bierwagen, K.A. Birngruber, M. Bosch, J. Breitlow, L.A. Cameron, J. Chalfoun, J.J. Chambers, C.L. Chen, E. Conde-Sousa, A.D. Corbett, F.P. Cordelieres, E.D. Nery, R. Dietzel, F. Eismann, E. Fazeli, A. Felscher, H. Fried, N. Gaudreault, W.I. Goh, T. Guilbert, R. Hadleigh, P. Hemmerich, G.A. Holst, M.S. Itano, C.B. Jaffe, H.K. Jambor, S.C. Jarvis, A. Keppler, D. Kirchenbuechler, M. Kirchner, N. Kobayashi, G. Krens, S. Kunis, J. Lacoste, M. Marcello, G.G. Martins, D.J. Metcalf, C.A. Mitchell, J. Moore, T. Mueller, M.S. Nelson, S. Ogg, S. Onami, A.L. Palmer, P. Paul-Gilloteaux, J.A. Pimentel, L. Plantard, S. Podder, E. Rexhepaj, A. Royon, M.A. Saari, D. Schapman, V. Schoonderwoert, B. Schroth-Diez, S. Schwartz, M. Shaw, M. Spitaler, M.T. Stoeckl, D. Sudar, J. Teillon, S. Terjung, R. Thuenauer, C.D. Wilms, G.D. Wright, R. Nitschke, Journal of Microscopy 284 (2021) 56–73. date_created: 2021-08-15T22:01:29Z date_published: 2021-08-11T00:00:00Z date_updated: 2023-08-11T10:30:40Z day: '11' department: - _id: Bio doi: 10.1111/jmi.13041 external_id: isi: - '000683702700001' intvolume: ' 284' isi: 1 issue: '1' language: - iso: eng main_file_link: - open_access: '1' url: https://doi.org/10.1111/jmi.13041 month: '08' oa: 1 oa_version: Published Version page: 56-73 publication: Journal of Microscopy publication_identifier: eissn: - 1365-2818 issn: - 0022-2720 publication_status: published publisher: Wiley quality_controlled: '1' scopus_import: '1' status: public title: 'QUAREP-LiMi: A community-driven initiative to establish guidelines for quality assessment and reproducibility for instruments and images in light microscopy' type: journal_article user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8 volume: 284 year: '2021' ...