Embryonic stem cells (ESCs) are a widely used model system to study cellular differentiation because of their pluripotent characteristics, and ESC differentiation is an epigenetic process. In an effort to identify a new epigenetic factor that is required for ESC differentiation, the function of SETD5 in ESCs was studied for this thesis. Results show that SETD5 is essential for retinoic acid (RA)-induced differentiation of mouse ESCs and for RA-induced expression of critical developmental genes (e.g., Hoxa1 and Hoxa2) and neuron-related genes (e.g., Nestin and Pax6). SETD5 was upregulated during ESC differentiation. Additional results demonstrated that SETD5 bound to RAR-α upon RA treatment and was recruited to a retinoic acid response element (RARE) for Hoxa1 and Hoxa2 activation. Methyltransferase assay using recombinant SETD5 and SETD5 complex showed that SETD5 was catalytically inactive, although it has a putative catalytic domain called SET. The transcription coactivator HCF1 was identified as a major SETD5-interacting protein. Depletion of HCF1 inhibited RA-induced ESC differentiation and RA-induced expression of SETD5-regulated genes. Chromatin immunoprecipitation assay provided evidence that HCF1 was localized to the SETD5-bound RARE region after RA treatment. These findings reveal a previously unknown ESC differentiation mechanism in which SETD5 facilitates mouse ESC differentiation by activating differentiation-specific genes via cooperation with HCF1.
In a related but independent study, we show that miR-221-3p and miR-221-5p, which are encoded by the miR-221 gene, are new anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by the epigenetic modifier and pluripotent factor PRMT7. Notably, both miR-221-3p and miR-221-5p can target the 3’ untranslated regions of the major pluripotent factors Oct4, Nanog and Sox2 to antagonize mouse ESC stemness while miR-221-5p additionally silences expression of its transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated miR-221 deletion and anti-sense miR-221 inhibitors inhibited spontaneous differentiation of PRMT7-depleted mouse ESCs. These results reveal that PRMT7-mediated repression of miR-221-3p and miR-221-5p is critical for maintaining mouse ESC stemness. Taken together, these two studies establish SETD5 and miR-221 as novel anti-stemness regulators that play a pro-differentiation role in ESC differentiation.
","abstract_html":"<p>Embryonic stem cells (ESCs) are a widely used model system to study cellular differentiation because of their pluripotent characteristics, and ESC differentiation is an epigenetic process. In an effort to identify a new epigenetic factor that is required for ESC differentiation, the function of SETD5 in ESCs was studied for this thesis. Results show that SETD5 is essential for retinoic acid (RA)-induced differentiation of mouse ESCs and for RA-induced expression of critical developmental genes (e.g., <em>Hoxa1</em> and <em>Hoxa2</em>) and neuron-related genes (e.g., <em>Nestin </em>and <em>Pax6</em>). SETD5 was upregulated during ESC differentiation. Additional results demonstrated that SETD5 bound to RAR-α upon RA treatment and was recruited to a<em> </em>retinoic acid response element (RARE) for <em>Hoxa1</em> and <em>Hoxa2 </em>activation. Methyltransferase assay using recombinant SETD5 and SETD5 complex showed that SETD5 was catalytically inactive, although it has a putative catalytic domain called SET. The transcription coactivator HCF1 was identified as a major SETD5-interacting protein. Depletion of HCF1 inhibited RA-induced ESC differentiation and RA-induced expression of SETD5-regulated genes. Chromatin immunoprecipitation assay provided evidence that HCF1 was localized to the SETD5-bound RARE region after RA treatment. These findings reveal a previously unknown ESC differentiation mechanism in which SETD5 facilitates mouse ESC differentiation by activating differentiation-specific genes via cooperation with HCF1.</p> <p>In a related but independent study, we show that miR-221-3p and miR-221-5p, which are encoded by the <em>miR-221</em> gene, are new anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by the epigenetic modifier and pluripotent factor PRMT7. Notably, both miR-221-3p and miR-221-5p can target the 3’ untranslated regions of the major pluripotent factors Oct4, Nanog and Sox2 to antagonize mouse ESC stemness while miR-221-5p additionally silences expression of its transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated <em>miR-221 </em>deletion and anti-sense <em>miR-221</em> inhibitors inhibited spontaneous differentiation of PRMT7-depleted mouse ESCs. These results reveal that PRMT7-mediated repression of miR-221-3p and miR-221-5p is critical for maintaining mouse ESC stemness. Taken together, these two studies establish SETD5 and <em>miR-221</em> as novel anti-stemness regulators that play a pro-differentiation role in ESC differentiation.</p>","abstract_has_math":false,"creators":["Chen, Tsai-Yu","https://orcid.org/0000-0001-6150-6361
"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation (PhD)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Min Gyu Lee","Xiaobing Shi","Jae-Il Park"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2017,"date_issued":"2017-12-01T08:00:00Z","date_published":"2017-12-01T08:00:00Z","updated_at":"2026-07-24T05:50:47Z","subjects":["ES cells","SETD5","miR-221","Retinoic acid","differentiation","HCF1","stemness","pluripotent factors","Medicine and Health Sciences","Molecular Biology","Other Cell and Developmental Biology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/817","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Min Gyu Lee","Xiaobing Shi","Jae-Il Park"]},{"key":"dc:creator","label":"Author","values":["Chen, Tsai-Yu","https://orcid.org/0000-0001-6150-6361
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In a related but independent study, we show that miR-221-3p and miR-221-5p, which are encoded by the miR-221 gene, are new anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by the epigenetic modifier and pluripotent factor PRMT7. Notably, both miR-221-3p and miR-221-5p can target the 3’ untranslated regions of the major pluripotent factors Oct4, Nanog and Sox2 to antagonize mouse ESC stemness while miR-221-5p additionally silences expression of its transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated miR-221 deletion and anti-sense miR-221 inhibitors inhibited spontaneous differentiation of PRMT7-depleted mouse ESCs. These results reveal that PRMT7-mediated repression of miR-221-3p and miR-221-5p is critical for maintaining mouse ESC stemness. Taken together, these two studies establish SETD5 and miR-221 as novel anti-stemness regulators that play a pro-differentiation role in ESC differentiation.
"]},{"key":"dc:title","label":"Title","values":["The Functions of Setd5 and Mir-221 In Embryonic Stem Cell Differentiation"]}]}],"canonical_facts":{"dc:contributor":["Min Gyu Lee","Xiaobing Shi","Jae-Il Park"],"dc:creator":["Chen, Tsai-Yu","https://orcid.org/0000-0001-6150-6361
"],"dc:date.available":["2018-11-30T08:00:00Z"],"dc:description.abstract":["Embryonic stem cells (ESCs) are a widely used model system to study cellular differentiation because of their pluripotent characteristics, and ESC differentiation is an epigenetic process. In an effort to identify a new epigenetic factor that is required for ESC differentiation, the function of SETD5 in ESCs was studied for this thesis. Results show that SETD5 is essential for retinoic acid (RA)-induced differentiation of mouse ESCs and for RA-induced expression of critical developmental genes (e.g., Hoxa1 and Hoxa2) and neuron-related genes (e.g., Nestin and Pax6). SETD5 was upregulated during ESC differentiation. Additional results demonstrated that SETD5 bound to RAR-α upon RA treatment and was recruited to a retinoic acid response element (RARE) for Hoxa1 and Hoxa2 activation. Methyltransferase assay using recombinant SETD5 and SETD5 complex showed that SETD5 was catalytically inactive, although it has a putative catalytic domain called SET. The transcription coactivator HCF1 was identified as a major SETD5-interacting protein. Depletion of HCF1 inhibited RA-induced ESC differentiation and RA-induced expression of SETD5-regulated genes. Chromatin immunoprecipitation assay provided evidence that HCF1 was localized to the SETD5-bound RARE region after RA treatment. These findings reveal a previously unknown ESC differentiation mechanism in which SETD5 facilitates mouse ESC differentiation by activating differentiation-specific genes via cooperation with HCF1.
In a related but independent study, we show that miR-221-3p and miR-221-5p, which are encoded by the miR-221 gene, are new anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by the epigenetic modifier and pluripotent factor PRMT7. Notably, both miR-221-3p and miR-221-5p can target the 3’ untranslated regions of the major pluripotent factors Oct4, Nanog and Sox2 to antagonize mouse ESC stemness while miR-221-5p additionally silences expression of its transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated miR-221 deletion and anti-sense miR-221 inhibitors inhibited spontaneous differentiation of PRMT7-depleted mouse ESCs. These results reveal that PRMT7-mediated repression of miR-221-3p and miR-221-5p is critical for maintaining mouse ESC stemness. Taken together, these two studies establish SETD5 and miR-221 as novel anti-stemness regulators that play a pro-differentiation role in ESC differentiation.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/817"],"dc:subject":["ES cells","SETD5","miR-221","Retinoic acid","differentiation","HCF1","stemness","pluripotent factors","Medicine and Health Sciences","Molecular Biology","Other Cell and Developmental Biology"],"dc:title":["The Functions of Setd5 and Mir-221 In Embryonic Stem Cell Differentiation"],"thesis:degree_level":["Dissertation (PhD)"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T05:50:47Z"}