{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/82383"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/82383","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Engineering Viruses for Gene Therapy: Isolating and Characterizing Murine Leukemia Virus With Improved Stability","abstract":"Inefficient gene delivery continues to be a primary hurdle facing gene therapy. Viruses offer the highest gene transfer capabilities but are not optimized as therapeutics. Applying directed evolution, we randomly mutated the entire genome of amphotropic murine leukemia virus (MLV) and selected for improved stability and infection at 37°C. After one round of mutagenesis and several rounds of selection, we isolated MLV variants with double the half-life of wild-type MLV. The improved stability of the mutant MLV leads to increased virus production, titer, and infection efficiency. Remarkably, a single mutation in the protease (PR), G119E, in the MLV gag-pro-pol is responsible for the enhanced stability. Thus, the variant MLV exhibits increased stability with various wild type envelope proteins, including amphotropic, ecotropic, 10A1, and VSV-G. Lastly, saturation mutagenesis at the site of the beneficial mutation identified MLV mutants with infectivity half-lives of ∼24 h at 37°C, nearly a 4-fold increase in infectivity half-life over the wild-type for this widely used gene therapy vector. Such engineered viral vectors may prove useful for future gene therapies.","abstract_html":"Inefficient gene delivery continues to be a primary hurdle facing gene therapy. Viruses offer the highest gene transfer capabilities but are not optimized as therapeutics. Applying directed evolution, we randomly mutated the entire genome of amphotropic murine leukemia virus (MLV) and selected for improved stability and infection at 37°C. After one round of mutagenesis and several rounds of selection, we isolated MLV variants with double the half-life of wild-type MLV. The improved stability of the mutant MLV leads to increased virus production, titer, and infection efficiency. Remarkably, a single mutation in the protease (PR), G119E, in the MLV gag-pro-pol is responsible for the enhanced stability. Thus, the variant MLV exhibits increased stability with various wild type envelope proteins, including amphotropic, ecotropic, 10A1, and VSV-G. Lastly, saturation mutagenesis at the site of the beneficial mutation identified MLV mutants with infectivity half-lives of ∼24 h at 37°C, nearly a 4-fold increase in infectivity half-life over the wild-type for this widely used gene therapy vector. Such engineered viral vectors may prove useful for future gene therapies.","abstract_has_math":false,"creators":["Vu, Halong Nguyen"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemical Engineering","degree_department":null,"school":null,"contributors":["Pack, Daniel W."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-09-25T20:43:26Z","date_published":"2015-09-25T20:43:26Z","updated_at":"2026-07-22T22:26:18Z","subjects":["Biology, Virology"],"languages":["eng"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(MiAaPQ)AAI3223740"],"render_values":[{"text":"(MiAaPQ)AAI3223740","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/82383","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Pack, Daniel W."]},{"key":"dc:creator","label":"Author","values":["Vu, Halong Nguyen"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2015-09-25T20:43:26Z","10000-01-01","2006"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemical Engineering"]},{"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":["Biology, Virology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/82383","(MiAaPQ)AAI3223740"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Inefficient gene delivery continues to be a primary hurdle facing gene therapy. Viruses offer the highest gene transfer capabilities but are not optimized as therapeutics. Applying directed evolution, we randomly mutated the entire genome of amphotropic murine leukemia virus (MLV) and selected for improved stability and infection at 37°C. After one round of mutagenesis and several rounds of selection, we isolated MLV variants with double the half-life of wild-type MLV. The improved stability of the mutant MLV leads to increased virus production, titer, and infection efficiency. Remarkably, a single mutation in the protease (PR), G119E, in the MLV gag-pro-pol is responsible for the enhanced stability. Thus, the variant MLV exhibits increased stability with various wild type envelope proteins, including amphotropic, ecotropic, 10A1, and VSV-G. Lastly, saturation mutagenesis at the site of the beneficial mutation identified MLV mutants with infectivity half-lives of ∼24 h at 37°C, nearly a 4-fold increase in infectivity half-life over the wild-type for this widely used gene therapy vector. Such engineered viral vectors may prove useful for future gene therapies.","Made available in DSpace on 2015-09-25T20:43:26Z (GMT). No. of bitstreams: 2 license.txt: 4848 bytes, checksum: 96035ab3f5e1c23cc7138a224ce498bd (MD5) 3223740.pdf: 3757077 bytes, checksum: f0dea7c6dbcacaffdb8a56fe719ee8ff (MD5) Previous issue date: 2006","Embargo set by: Seth Robbins for item 83664 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","130 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2006."]},{"key":"dc:title","label":"Title","values":["Engineering Viruses for Gene Therapy: Isolating and Characterizing Murine Leukemia Virus With Improved Stability"]}]}],"canonical_facts":{"dc:contributor":["Pack, Daniel W."],"dc:creator":["Vu, Halong Nguyen"],"dc:date":["2015-09-25T20:43:26Z","10000-01-01","2006"],"dc:description":["Inefficient gene delivery continues to be a primary hurdle facing gene therapy. Viruses offer the highest gene transfer capabilities but are not optimized as therapeutics. Applying directed evolution, we randomly mutated the entire genome of amphotropic murine leukemia virus (MLV) and selected for improved stability and infection at 37°C. After one round of mutagenesis and several rounds of selection, we isolated MLV variants with double the half-life of wild-type MLV. The improved stability of the mutant MLV leads to increased virus production, titer, and infection efficiency. Remarkably, a single mutation in the protease (PR), G119E, in the MLV gag-pro-pol is responsible for the enhanced stability. Thus, the variant MLV exhibits increased stability with various wild type envelope proteins, including amphotropic, ecotropic, 10A1, and VSV-G. Lastly, saturation mutagenesis at the site of the beneficial mutation identified MLV mutants with infectivity half-lives of ∼24 h at 37°C, nearly a 4-fold increase in infectivity half-life over the wild-type for this widely used gene therapy vector. Such engineered viral vectors may prove useful for future gene therapies.","Made available in DSpace on 2015-09-25T20:43:26Z (GMT). No. of bitstreams: 2 license.txt: 4848 bytes, checksum: 96035ab3f5e1c23cc7138a224ce498bd (MD5) 3223740.pdf: 3757077 bytes, checksum: f0dea7c6dbcacaffdb8a56fe719ee8ff (MD5) Previous issue date: 2006","Embargo set by: Seth Robbins for item 83664 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","130 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2006."],"dc:identifier":["http://hdl.handle.net/2142/82383","(MiAaPQ)AAI3223740"],"dc:language":["eng"],"dc:subject":["Biology, Virology"],"dc:title":["Engineering Viruses for Gene Therapy: Isolating and Characterizing Murine Leukemia Virus With Improved Stability"],"dc:type":["text"],"thesis:degree_discipline":["Chemical Engineering"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:18Z"}