{"id":{"repo_id":"penn","oai_identifier":"oai:repository.upenn.edu:20.500.14332/28506"},"canonical_url":"https://search.dev.ndltd.org/etd/penn/oai:repository.upenn.edu:20.500.14332/28506","repository":{"repo_id":"penn","name":"University of Pennsylvania","base_url":"https://repository.upenn.edu/server/oai/request"},"display":{"title":"The Cellular Mirna, Mir-190, is Upregulated in Type I Ebv Latency by Ebers and Modulates Cellular Mrnas involved in Cell Survival and Viral Reactivation","abstract":"Epstein-Barr Virus (EBV) is a highly prevalent human pathogen infecting over 90% of the population. Much of the success of the virus is attributed to its ability to maintain latency through different programs in host cells. MicroRNAs (miRNA) are small, non-coding RNAs capable of post-transcriptionally regulating mRNA expression. A microarray comparison of EBV type I latency and type III latency infected cells yielded evidence of differential cellular microRNA expression. I hypothesized that one of these differentially upregulated type I latency miRNAs, miR-190, is important in maintenance of latency I, and miR-190 upregulation is due to viral gene expression. Lentiviral overexpression systems were used to overexpress miR-190 and a microarray of gene expression revealed candidate miR-190 targets, including: TP53INP1 and NR4A3. The modulation of these targets by miR-190 was confirmed through evaluating mRNA and protein level changes in the presence or absence of miR-190. In the case of TP53INP1, a 3’UTR target site was identified through mutagenesis. The effect of miR-190 expression was evaluated for markers of cell cycle and cell death by flow cytometry, western blot and RT-PCR. Measures of viral reactivation were lowered in the presence of miR-190 after induction by anti-IgG stimulation. I also observed upregulation of miR-190/Talin2 promoter activity or miR-190 expression in the presence of EBERs, Epstein-Barr encoded RNAs. Interestingly, a panel of type I latency cell lines had higher EBER1 expression compared to their type III latency counterparts. Work by others has indicated that EBERs activate the double-stranded RNA (dsRNA) sensor, retinoic acid-inducible gene 1 (RIG-I). Transiently expressed, constitutively activated RIG-I induced miR-190 expression and promoter activity. Knockdown of RIG-I in the type I latency cells yielded lowered miR-190 expression levels. To investigate how miR-190 is upregulated I generated miR-190/Talin2 promoter reporters that lacked YinYang1 (YY1) and Nuclear factor-κB (NF-kB) binding motifs. In the presence of EBERs, promoters with these deleted binding motifs had lowered activation compared to the full miR-190/TLN2 promoter. This work describes a mechanism by which EBERs upregulate a cellular miRNA, miR-190, which aids in type I latency preservation by preventing apoptosis, promoting cell cycle and maintaining virus in its latent state.","abstract_html":"Epstein-Barr Virus (EBV) is a highly prevalent human pathogen infecting over 90% of the population. Much of the success of the virus is attributed to its ability to maintain latency through different programs in host cells. MicroRNAs (miRNA) are small, non-coding RNAs capable of post-transcriptionally regulating mRNA expression. A microarray comparison of EBV type I latency and type III latency infected cells yielded evidence of differential cellular microRNA expression. I hypothesized that one of these differentially upregulated type I latency miRNAs, miR-190, is important in maintenance of latency I, and miR-190 upregulation is due to viral gene expression. Lentiviral overexpression systems were used to overexpress miR-190 and a microarray of gene expression revealed candidate miR-190 targets, including: TP53INP1 and NR4A3. The modulation of these targets by miR-190 was confirmed through evaluating mRNA and protein level changes in the presence or absence of miR-190. In the case of TP53INP1, a 3’UTR target site was identified through mutagenesis. The effect of miR-190 expression was evaluated for markers of cell cycle and cell death by flow cytometry, western blot and RT-PCR. Measures of viral reactivation were lowered in the presence of miR-190 after induction by anti-IgG stimulation. I also observed upregulation of miR-190/Talin2 promoter activity or miR-190 expression in the presence of EBERs, Epstein-Barr encoded RNAs. Interestingly, a panel of type I latency cell lines had higher EBER1 expression compared to their type III latency counterparts. Work by others has indicated that EBERs activate the double-stranded RNA (dsRNA) sensor, retinoic acid-inducible gene 1 (RIG-I). Transiently expressed, constitutively activated RIG-I induced miR-190 expression and promoter activity. Knockdown of RIG-I in the type I latency cells yielded lowered miR-190 expression levels. To investigate how miR-190 is upregulated I generated miR-190/Talin2 promoter reporters that lacked YinYang1 (YY1) and Nuclear factor-κB (NF-kB) binding motifs. In the presence of EBERs, promoters with these deleted binding motifs had lowered activation compared to the full miR-190/TLN2 promoter. This work describes a mechanism by which EBERs upregulate a cellular miRNA, miR-190, which aids in type I latency preservation by preventing apoptosis, promoting cell cycle and maintaining virus in its latent state.","abstract_has_math":false,"creators":["Cramer, Elizabeth Mary"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Yan Yuan"],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-01-01","date_published":"2015-01-01","updated_at":"2026-07-24T03:47:17Z","subjects":[],"languages":["en"],"rights":["Elizabeth Mary Cramer"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://repository.upenn.edu/handle/20.500.14332/28506","outbound_label":"Repository record","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Yan Yuan"]},{"key":"dc:creator","label":"Author","values":["Cramer, Elizabeth Mary"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2023-05-17T15:54:08.000"]},{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2023-05-22T16:43:18Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2015-07-22T00:00:00Z"]},{"key":"dc:date.issued","label":"Date","values":["2015-01-01"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation/Thesis"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Elizabeth Mary Cramer"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://repository.upenn.edu/handle/20.500.14332/28506"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Epstein-Barr Virus (EBV) is a highly prevalent human pathogen infecting over 90% of the population. Much of the success of the virus is attributed to its ability to maintain latency through different programs in host cells. MicroRNAs (miRNA) are small, non-coding RNAs capable of post-transcriptionally regulating mRNA expression. A microarray comparison of EBV type I latency and type III latency infected cells yielded evidence of differential cellular microRNA expression. I hypothesized that one of these differentially upregulated type I latency miRNAs, miR-190, is important in maintenance of latency I, and miR-190 upregulation is due to viral gene expression. Lentiviral overexpression systems were used to overexpress miR-190 and a microarray of gene expression revealed candidate miR-190 targets, including: TP53INP1 and NR4A3. The modulation of these targets by miR-190 was confirmed through evaluating mRNA and protein level changes in the presence or absence of miR-190. In the case of TP53INP1, a 3’UTR target site was identified through mutagenesis. The effect of miR-190 expression was evaluated for markers of cell cycle and cell death by flow cytometry, western blot and RT-PCR. Measures of viral reactivation were lowered in the presence of miR-190 after induction by anti-IgG stimulation. I also observed upregulation of miR-190/Talin2 promoter activity or miR-190 expression in the presence of EBERs, Epstein-Barr encoded RNAs. Interestingly, a panel of type I latency cell lines had higher EBER1 expression compared to their type III latency counterparts. Work by others has indicated that EBERs activate the double-stranded RNA (dsRNA) sensor, retinoic acid-inducible gene 1 (RIG-I). Transiently expressed, constitutively activated RIG-I induced miR-190 expression and promoter activity. Knockdown of RIG-I in the type I latency cells yielded lowered miR-190 expression levels. To investigate how miR-190 is upregulated I generated miR-190/Talin2 promoter reporters that lacked YinYang1 (YY1) and Nuclear factor-κB (NF-kB) binding motifs. In the presence of EBERs, promoters with these deleted binding motifs had lowered activation compared to the full miR-190/TLN2 promoter. This work describes a mechanism by which EBERs upregulate a cellular miRNA, miR-190, which aids in type I latency preservation by preventing apoptosis, promoting cell cycle and maintaining virus in its latent state."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Doctor of Philosophy (PhD)"]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["The Cellular Mirna, Mir-190, is Upregulated in Type I Ebv Latency by Ebers and Modulates Cellular Mrnas involved in Cell Survival and Viral Reactivation"]}]}],"canonical_facts":{"dc:contributor.advisor":["Yan Yuan"],"dc:creator":["Cramer, Elizabeth Mary"],"dc:date":["2023-05-17T15:54:08.000"],"dc:date.accessioned":["2023-05-22T16:43:18Z"],"dc:date.available":["2015-07-22T00:00:00Z"],"dc:date.issued":["2015-01-01"],"dc:description.abstract":["Epstein-Barr Virus (EBV) is a highly prevalent human pathogen infecting over 90% of the population. Much of the success of the virus is attributed to its ability to maintain latency through different programs in host cells. MicroRNAs (miRNA) are small, non-coding RNAs capable of post-transcriptionally regulating mRNA expression. A microarray comparison of EBV type I latency and type III latency infected cells yielded evidence of differential cellular microRNA expression. I hypothesized that one of these differentially upregulated type I latency miRNAs, miR-190, is important in maintenance of latency I, and miR-190 upregulation is due to viral gene expression. Lentiviral overexpression systems were used to overexpress miR-190 and a microarray of gene expression revealed candidate miR-190 targets, including: TP53INP1 and NR4A3. The modulation of these targets by miR-190 was confirmed through evaluating mRNA and protein level changes in the presence or absence of miR-190. In the case of TP53INP1, a 3’UTR target site was identified through mutagenesis. The effect of miR-190 expression was evaluated for markers of cell cycle and cell death by flow cytometry, western blot and RT-PCR. Measures of viral reactivation were lowered in the presence of miR-190 after induction by anti-IgG stimulation. I also observed upregulation of miR-190/Talin2 promoter activity or miR-190 expression in the presence of EBERs, Epstein-Barr encoded RNAs. Interestingly, a panel of type I latency cell lines had higher EBER1 expression compared to their type III latency counterparts. Work by others has indicated that EBERs activate the double-stranded RNA (dsRNA) sensor, retinoic acid-inducible gene 1 (RIG-I). Transiently expressed, constitutively activated RIG-I induced miR-190 expression and promoter activity. Knockdown of RIG-I in the type I latency cells yielded lowered miR-190 expression levels. To investigate how miR-190 is upregulated I generated miR-190/Talin2 promoter reporters that lacked YinYang1 (YY1) and Nuclear factor-κB (NF-kB) binding motifs. In the presence of EBERs, promoters with these deleted binding motifs had lowered activation compared to the full miR-190/TLN2 promoter. This work describes a mechanism by which EBERs upregulate a cellular miRNA, miR-190, which aids in type I latency preservation by preventing apoptosis, promoting cell cycle and maintaining virus in its latent state."],"dc:description.degree":["Doctor of Philosophy (PhD)"],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["https://repository.upenn.edu/handle/20.500.14332/28506"],"dc:language":["en"],"dc:rights":["Elizabeth Mary Cramer"],"dc:title":["The Cellular Mirna, Mir-190, is Upregulated in Type I Ebv Latency by Ebers and Modulates Cellular Mrnas involved in Cell Survival and Viral Reactivation"],"dc:type":["Dissertation/Thesis"]},"updated_at":"2026-07-24T03:47:17Z"}