{"id":{"repo_id":"ku","oai_identifier":"oai:kuscholarworks.ku.edu:1808/36672"},"canonical_url":"https://search.dev.ndltd.org/etd/ku/oai:kuscholarworks.ku.edu:1808/36672","repository":{"repo_id":"ku","name":"University of Kansas","base_url":"https://kuscholarworks.ku.edu/server/oai/request"},"display":{"title":"Development of a tool for tracking H3K27 tri-methylation","abstract":"H3K27 trimethylation is a post-translational modification known to play a role in faithful chromosome segregation via its interaction with TopoIIα. The ability to observe the spatiotemporal dynamics of this modification and how it interacts with TopoIIα has thus far been a challenge. Here, we have utilized a method involving an inducible fluorescent modification-specific intracellular antibody (mintbody) that reversibly binds to post-translational modifications to tag the H3K27 trimethylation. Four cell lines were created using a mintbody including super folded GFP that can be induced with treatment with doxycycline. Different numbers of guide RNAs were also used in an attempt to increase CRISPR/Cas9 efficiency. These cell lines were screened and analyzed for future use in live cell imaging and in vivo experiments concerning the interplay of H3K27 trimethylation, TopoIIα, and chromosome segregation.","abstract_html":"H3K27 trimethylation is a post-translational modification known to play a role in faithful chromosome segregation via its interaction with TopoIIα. The ability to observe the spatiotemporal dynamics of this modification and how it interacts with TopoIIα has thus far been a challenge. Here, we have utilized a method involving an inducible fluorescent modification-specific intracellular antibody (mintbody) that reversibly binds to post-translational modifications to tag the H3K27 trimethylation. Four cell lines were created using a mintbody including super folded GFP that can be induced with treatment with doxycycline. Different numbers of guide RNAs were also used in an attempt to increase CRISPR/Cas9 efficiency. These cell lines were screened and analyzed for future use in live cell imaging and in vivo experiments concerning the interplay of H3K27 trimethylation, TopoIIα, and chromosome segregation.","abstract_has_math":false,"creators":["Bertolino, Morgan"],"institution":"University of Kansas","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Azuma, Yoshiaki"],"committee_chairs":[],"committee_members":[],"year":2024,"date_issued":"2024-12-31","date_published":"2024-12-31","updated_at":"2026-07-24T02:46:17Z","subjects":["Cellular biology","Molecular biology","CRISPR","H3K27me3","histone posttranslational modifications","mintbody"],"languages":["en"],"rights":["This item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author."],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["http://dissertations.umi.com/ku:19743"],"render_values":[{"text":"http://dissertations.umi.com/ku:19743","href":"http://dissertations.umi.com/ku:19743","code":true}]}]},"links":{"outbound_url":"https://hdl.handle.net/1808/36672","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Azuma, Yoshiaki"]},{"key":"dc:creator","label":"Author","values":["Bertolino, Morgan"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2026-04-12T19:43:44Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2026-04-12T19:43:44Z"]},{"key":"dc:date.issued","label":"Date","values":["2024-12-31"]},{"key":"dc:publisher","label":"Institution","values":["University of Kansas"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Cellular biology","Molecular biology","CRISPR","H3K27me3","histone posttranslational modifications","mintbody"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["This item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["http://dissertations.umi.com/ku:19743"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/1808/36672"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["H3K27 trimethylation is a post-translational modification known to play a role in faithful chromosome segregation via its interaction with TopoIIα. 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These cell lines were screened and analyzed for future use in live cell imaging and in vivo experiments concerning the interplay of H3K27 trimethylation, TopoIIα, and chromosome segregation."]},{"key":"dc:title","label":"Title","values":["Development of a tool for tracking H3K27 tri-methylation"]}]}],"canonical_facts":{"dc:contributor.advisor":["Azuma, Yoshiaki"],"dc:creator":["Bertolino, Morgan"],"dc:date.accessioned":["2026-04-12T19:43:44Z"],"dc:date.available":["2026-04-12T19:43:44Z"],"dc:date.issued":["2024-12-31"],"dc:description.abstract":["H3K27 trimethylation is a post-translational modification known to play a role in faithful chromosome segregation via its interaction with TopoIIα. The ability to observe the spatiotemporal dynamics of this modification and how it interacts with TopoIIα has thus far been a challenge. Here, we have utilized a method involving an inducible fluorescent modification-specific intracellular antibody (mintbody) that reversibly binds to post-translational modifications to tag the H3K27 trimethylation. Four cell lines were created using a mintbody including super folded GFP that can be induced with treatment with doxycycline. Different numbers of guide RNAs were also used in an attempt to increase CRISPR/Cas9 efficiency. 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