{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/105909"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/105909","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Creating a versatile toolkit for transgene expression using BACS and for dissecting large-scale chromatin organization","abstract":"In eukaryotes, the genetic material, DNA, is highly compacted with histone proteins to form chromatin in interphase nuclei. Both the higher levels of chromatin folding and the spatial organization of the chromatin, referred to as large-scale chromatin organization, have been shown to correlate with transcriptional activity. One example suggesting transcriptional regulation by large-scale chromatin organization is position effects and position effect variegations observed in transgene expressions, which, as well as other epigenetic silencing mechanisms, have been obstacles to achieving predictable and stable transgene expression. Molecular dissections of the determinants regulating large-scale chromatin organization would help to elucidate the real relationship between large-scale chromatin organization and transcriptional regulation, yet are difficult due to the complexity of the mammalian genome. Bacterial artificial chromosomes (BACs), containing 100-300 kb mammalian genomic regions have been shown to recapitulate the expression level and nuclear localization of the corresponding genomic regions, and to protect embedded reporter mini-genes from epigenetic silencing. Here in Chapter 2 we show that BACs could provide a versatile platform for achieving reproducible, stable simultaneous expression of multiple transgenes maintained either as episomes or stably integrated copies. Moreover, in Chapter 3 we show that BACs could be used as a powerful model system for dissecting mechanisms regulating large-scale chromatin organization, by demonstrating distinctive large-scale chromatin conformations formed by BAC transgene arrays and results indicating separation of large-scale chromatin compaction, nuclear localization and transcriptional activities.","abstract_html":"In eukaryotes, the genetic material, DNA, is highly compacted with histone proteins to form chromatin in interphase nuclei. Both the higher levels of chromatin folding and the spatial organization of the chromatin, referred to as large-scale chromatin organization, have been shown to correlate with transcriptional activity. One example suggesting transcriptional regulation by large-scale chromatin organization is position effects and position effect variegations observed in transgene expressions, which, as well as other epigenetic silencing mechanisms, have been obstacles to achieving predictable and stable transgene expression. Molecular dissections of the determinants regulating large-scale chromatin organization would help to elucidate the real relationship between large-scale chromatin organization and transcriptional regulation, yet are difficult due to the complexity of the mammalian genome. Bacterial artificial chromosomes (BACs), containing 100-300 kb mammalian genomic regions have been shown to recapitulate the expression level and nuclear localization of the corresponding genomic regions, and to protect embedded reporter mini-genes from epigenetic silencing. Here in Chapter 2 we show that BACs could provide a versatile platform for achieving reproducible, stable simultaneous expression of multiple transgenes maintained either as episomes or stably integrated copies. Moreover, in Chapter 3 we show that BACs could be used as a powerful model system for dissecting mechanisms regulating large-scale chromatin organization, by demonstrating distinctive large-scale chromatin conformations formed by BAC transgene arrays and results indicating separation of large-scale chromatin compaction, nuclear localization and transcriptional activities.","abstract_has_math":false,"creators":["Zhao, Binhui"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Cell and Developmental Biology","degree_department":null,"school":null,"contributors":["Belmont, Andrew S","Prasanth, Supriya","Stubbs, Lisa J","Freeman, Brian C","Zhao, Humin"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-11-26T20:59:34Z","date_published":"2019-11-26T20:59:34Z","updated_at":"2026-07-22T22:24:45Z","subjects":["transgene expression","episomes","gene amplification","human artificial chromosomes","DNA synthesis","chromatin conformation","transcription regulation","CRISPR/Cas"],"languages":["en"],"rights":["Copyright 2019 Binhui Zhao"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/105909","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Belmont, Andrew S","Prasanth, Supriya","Stubbs, Lisa J","Freeman, Brian C","Zhao, Humin"]},{"key":"dc:creator","label":"Author","values":["Zhao, Binhui"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-11-26T20:59:34Z","2021-11-27T10:15:09Z","2019-07-09","2019-08"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Cell and Developmental Biology"]},{"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":["transgene expression","episomes","gene amplification","human artificial chromosomes","DNA synthesis","chromatin conformation","transcription regulation","CRISPR/Cas"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2019 Binhui Zhao"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/105909"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["In eukaryotes, the genetic material, DNA, is highly compacted with histone proteins to form chromatin in interphase nuclei. Both the higher levels of chromatin folding and the spatial organization of the chromatin, referred to as large-scale chromatin organization, have been shown to correlate with transcriptional activity. One example suggesting transcriptional regulation by large-scale chromatin organization is position effects and position effect variegations observed in transgene expressions, which, as well as other epigenetic silencing mechanisms, have been obstacles to achieving predictable and stable transgene expression. Molecular dissections of the determinants regulating large-scale chromatin organization would help to elucidate the real relationship between large-scale chromatin organization and transcriptional regulation, yet are difficult due to the complexity of the mammalian genome. Bacterial artificial chromosomes (BACs), containing 100-300 kb mammalian genomic regions have been shown to recapitulate the expression level and nuclear localization of the corresponding genomic regions, and to protect embedded reporter mini-genes from epigenetic silencing. Here in Chapter 2 we show that BACs could provide a versatile platform for achieving reproducible, stable simultaneous expression of multiple transgenes maintained either as episomes or stably integrated copies. Moreover, in Chapter 3 we show that BACs could be used as a powerful model system for dissecting mechanisms regulating large-scale chromatin organization, by demonstrating distinctive large-scale chromatin conformations formed by BAC transgene arrays and results indicating separation of large-scale chromatin compaction, nuclear localization and transcriptional activities.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-08-01","The student, Binhui Zhao, accepted the attached license on 2019-07-08 at 10:18.","The student, Binhui Zhao, submitted this Dissertation for approval on 2019-07-08 at 10:21.","This Dissertation was approved for publication on 2019-07-09 at 11:09.","DSpace SAF Submission Ingestion Package generated from Vireo submission #14187 on 2019-11-26 at 14:01:40","Made available in DSpace on 2019-11-26T20:59:34Z (GMT). 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Both the higher levels of chromatin folding and the spatial organization of the chromatin, referred to as large-scale chromatin organization, have been shown to correlate with transcriptional activity. One example suggesting transcriptional regulation by large-scale chromatin organization is position effects and position effect variegations observed in transgene expressions, which, as well as other epigenetic silencing mechanisms, have been obstacles to achieving predictable and stable transgene expression. Molecular dissections of the determinants regulating large-scale chromatin organization would help to elucidate the real relationship between large-scale chromatin organization and transcriptional regulation, yet are difficult due to the complexity of the mammalian genome. Bacterial artificial chromosomes (BACs), containing 100-300 kb mammalian genomic regions have been shown to recapitulate the expression level and nuclear localization of the corresponding genomic regions, and to protect embedded reporter mini-genes from epigenetic silencing. Here in Chapter 2 we show that BACs could provide a versatile platform for achieving reproducible, stable simultaneous expression of multiple transgenes maintained either as episomes or stably integrated copies. Moreover, in Chapter 3 we show that BACs could be used as a powerful model system for dissecting mechanisms regulating large-scale chromatin organization, by demonstrating distinctive large-scale chromatin conformations formed by BAC transgene arrays and results indicating separation of large-scale chromatin compaction, nuclear localization and transcriptional activities.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-08-01","The student, Binhui Zhao, accepted the attached license on 2019-07-08 at 10:18.","The student, Binhui Zhao, submitted this Dissertation for approval on 2019-07-08 at 10:21.","This Dissertation was approved for publication on 2019-07-09 at 11:09.","DSpace SAF Submission Ingestion Package generated from Vireo submission #14187 on 2019-11-26 at 14:01:40","Made available in DSpace on 2019-11-26T20:59:34Z (GMT). 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