2026-09-20T07:40:20Zhttps://digital.lib.washington.edu/server/oai/request

oai:digital.lib.washington.edu:1773/497262026-02-15T21:17:23Zcom_1773_4888col_1773_4943

Emerman, Michael Montoya, Vanessa Renae 2023-01-21T05:04:49Z 2023-01-21T05:04:49Z 2023-01-21 2022 Montoya_washington_0250E_24969.pdf http://hdl.handle.net/1773/49726 Thesis (Ph.D.)--University of Washington, 2022 In this thesis, I developed a method called HIV-CRISPR screening at genome-wide scale, and then at a smaller, targeted scale, to uncover novel genes that may act as HIV-1 dependency factors. By using a screening technique which assesses enrichment or depletion of guide based on HIV-1 release from infected cells, I was able to assess the role of host factors across the entire life cycle of HIV in a T lymphocyte cell line. Host dependency factors for HIV-1 were uncovered across multiple strains, from different clades and with different co-receptor tropisms. Nearly all of the host dependency factors were validated in subsequent follow-up studies and there was a direct relationship between the scoring of a dependency factor in the screen and its effect on HIV replication in validation studies. Work from this thesis thus identifies at least several dozen novel HIV dependency factors across multiple pathways. application/pdf en_US Supplementary Table 1. MAGeCK gene scores.xlsx; spreadsheet; MAGeCK gene scores for each HIV-CRISPR screen. Supplementary Table 2. Complete list of the HIVDEP genes and sgRNA sequences.xlsx; spreadsheet; HIVDEP CRISPR guide library information. Supplementary Table 3. z scores for each HIVDEP screen and Literature Review top genes.xlsx; spreadsheet; z score analysis for each HIVDEP screen and literature review for top hits. none CRISPR/Cas9 screen Dependency factors genome-wide screen HIV Human Immunodeficiency Virus transmitted-founder virus Virology Cellular biology Microbiology Molecular and cellular biology Genome scale identification of HIV Dependency factors across multiple HIV-1 strains Thesis Open Access open.access