2026-09-22T23:35:34Z https://www.ideals.illinois.edu/oai-pmh

oai:www.ideals.illinois.edu:2142/82383 2023-07-11 col_2142_5131 col_2142_11615 com_2142_5130 com_2142_9130 com_2142_8903

Pack, Daniel W. Vu, Halong Nguyen 2015-09-25T20:43:26Z 2015-09-25T20:43:26Z 10000-01-01 2006 2006 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. http://hdl.handle.net/2142/82383 (MiAaPQ)AAI3223740 eng Biology, Virology Engineering Viruses for Gene Therapy: Isolating and Characterizing Murine Leukemia Virus With Improved Stability text Chemical Engineering Chemical Engineering University of Illinois at Urbana-Champaign Dissertation Ph.D.