{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/108280"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/108280","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Optogenetic regulation of protein activity in live cell","abstract":"Signaling pathways extensively crosstalk among each other and result in different cellular phenotypes depending on the dynamic profile of protein activity. Conventional genetic and pharmacological approaches such as gene overexpression, use of growth factors or inhibitors have helped us delineate interaction maps of signaling components, however these techniques provide limited means to determine contribution of a target protein for specific cellular phenotype. Therefore, to find out the molecular mechanism for a cellular outcome, there is an urgent need for a tool that can specifically activate or inactivate a protein of interest and study it’s role towards a specific cell fate. Optogenetic techniques utilize light to control protein functions with high spatial and temporal resolution. Here, I First present a generalizable light modulated protein stabilization system (GLIMPSe) that enables target-independent optogenetic control of protein activities and minimizes the systematic variation embedded within different photoactivatable proteins. GLIMPSe was applied to control light-mediated post-translational stabilization of two distinct classes of proteins, phosphatase and kinase with rapid kinetics response. Second, I discuss role of speckle-type POZ protein (SPOP) in cell differentiation inhibition in PC12 and primary rat hippocampus neuron cell. Next, I combined the GLIMPSe system with the Sufu protein, a SPOP phenocopy and sonic hedgehog (Shh) signaling pathway inhibitor and generated the GLIMPSe-Sufu system for optogenetic inhibition of Shh signaling pathway. Finally, I review the molecular machinery of cargo trafficking with emphasis on new optogenetic and optochemical experimental strategies that enable direct modulation of cargo trafficking in live cells.","abstract_html":"Signaling pathways extensively crosstalk among each other and result in different cellular phenotypes depending on the dynamic profile of protein activity. Conventional genetic and pharmacological approaches such as gene overexpression, use of growth factors or inhibitors have helped us delineate interaction maps of signaling components, however these techniques provide limited means to determine contribution of a target protein for specific cellular phenotype. Therefore, to find out the molecular mechanism for a cellular outcome, there is an urgent need for a tool that can specifically activate or inactivate a protein of interest and study it’s role towards a specific cell fate. Optogenetic techniques utilize light to control protein functions with high spatial and temporal resolution. Here, I First present a generalizable light modulated protein stabilization system (GLIMPSe) that enables target-independent optogenetic control of protein activities and minimizes the systematic variation embedded within different photoactivatable proteins. GLIMPSe was applied to control light-mediated post-translational stabilization of two distinct classes of proteins, phosphatase and kinase with rapid kinetics response. Second, I discuss role of speckle-type POZ protein (SPOP) in cell differentiation inhibition in PC12 and primary rat hippocampus neuron cell. Next, I combined the GLIMPSe system with the Sufu protein, a SPOP phenocopy and sonic hedgehog (Shh) signaling pathway inhibitor and generated the GLIMPSe-Sufu system for optogenetic inhibition of Shh signaling pathway. Finally, I review the molecular machinery of cargo trafficking with emphasis on new optogenetic and optochemical experimental strategies that enable direct modulation of cargo trafficking in live cells.","abstract_has_math":false,"creators":["Mondal, Payel"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Biochemistry","degree_department":null,"school":null,"contributors":["Zhang, Kai","Chen, Lin-Feng","Chen, Jie","Kalsotra, Auinash"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-08-27T00:50:04Z","date_published":"2020-08-27T00:50:04Z","updated_at":"2026-07-22T22:24:48Z","subjects":["optogenetics","GLIMPSe","degron","protein degradation","MKP3","CA MEK","BRD4","SPOP","PC12","Sonic Hedgehog","Sufu","Gli","Cargo Trafficking"],"languages":["en"],"rights":["Copyright 2020 Payel Mondal"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/108280","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Zhang, Kai","Chen, Lin-Feng","Chen, Jie","Kalsotra, Auinash"]},{"key":"dc:creator","label":"Author","values":["Mondal, Payel"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2020-08-27T00:50:04Z","2022-08-27T00:51:40Z","2020-05-07","2020-05"]},{"key":"dc:type","label":"Dc Type","values":["text","Thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["optogenetics","GLIMPSe","degron","protein degradation","MKP3","CA MEK","BRD4","SPOP","PC12","Sonic Hedgehog","Sufu","Gli","Cargo Trafficking"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2020 Payel Mondal"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/108280"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Signaling pathways extensively crosstalk among each other and result in different cellular phenotypes depending on the dynamic profile of protein activity. Conventional genetic and pharmacological approaches such as gene overexpression, use of growth factors or inhibitors have helped us delineate interaction maps of signaling components, however these techniques provide limited means to determine contribution of a target protein for specific cellular phenotype. Therefore, to find out the molecular mechanism for a cellular outcome, there is an urgent need for a tool that can specifically activate or inactivate a protein of interest and study it’s role towards a specific cell fate. Optogenetic techniques utilize light to control protein functions with high spatial and temporal resolution. Here, I First present a generalizable light modulated protein stabilization system (GLIMPSe) that enables target-independent optogenetic control of protein activities and minimizes the systematic variation embedded within different photoactivatable proteins. GLIMPSe was applied to control light-mediated post-translational stabilization of two distinct classes of proteins, phosphatase and kinase with rapid kinetics response. Second, I discuss role of speckle-type POZ protein (SPOP) in cell differentiation inhibition in PC12 and primary rat hippocampus neuron cell. Next, I combined the GLIMPSe system with the Sufu protein, a SPOP phenocopy and sonic hedgehog (Shh) signaling pathway inhibitor and generated the GLIMPSe-Sufu system for optogenetic inhibition of Shh signaling pathway. Finally, I review the molecular machinery of cargo trafficking with emphasis on new optogenetic and optochemical experimental strategies that enable direct modulation of cargo trafficking in live cells.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2022-05-01","The student, Payel Mondal, accepted the attached license on 2020-05-01 at 14:55.","The student, Payel Mondal, submitted this Dissertation for approval on 2020-05-01 at 15:01.","This Dissertation was approved for publication on 2020-05-07 at 14:44.","DSpace SAF Submission Ingestion Package generated from Vireo submission #15084 on 2020-08-25 at 17:41:15","Made available in DSpace on 2020-08-27T00:50:04Z (GMT). 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Conventional genetic and pharmacological approaches such as gene overexpression, use of growth factors or inhibitors have helped us delineate interaction maps of signaling components, however these techniques provide limited means to determine contribution of a target protein for specific cellular phenotype. Therefore, to find out the molecular mechanism for a cellular outcome, there is an urgent need for a tool that can specifically activate or inactivate a protein of interest and study it’s role towards a specific cell fate. Optogenetic techniques utilize light to control protein functions with high spatial and temporal resolution. Here, I First present a generalizable light modulated protein stabilization system (GLIMPSe) that enables target-independent optogenetic control of protein activities and minimizes the systematic variation embedded within different photoactivatable proteins. GLIMPSe was applied to control light-mediated post-translational stabilization of two distinct classes of proteins, phosphatase and kinase with rapid kinetics response. Second, I discuss role of speckle-type POZ protein (SPOP) in cell differentiation inhibition in PC12 and primary rat hippocampus neuron cell. Next, I combined the GLIMPSe system with the Sufu protein, a SPOP phenocopy and sonic hedgehog (Shh) signaling pathway inhibitor and generated the GLIMPSe-Sufu system for optogenetic inhibition of Shh signaling pathway. Finally, I review the molecular machinery of cargo trafficking with emphasis on new optogenetic and optochemical experimental strategies that enable direct modulation of cargo trafficking in live cells.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2022-05-01","The student, Payel Mondal, accepted the attached license on 2020-05-01 at 14:55.","The student, Payel Mondal, submitted this Dissertation for approval on 2020-05-01 at 15:01.","This Dissertation was approved for publication on 2020-05-07 at 14:44.","DSpace SAF Submission Ingestion Package generated from Vireo submission #15084 on 2020-08-25 at 17:41:15","Made available in DSpace on 2020-08-27T00:50:04Z (GMT). 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