Identication and characterization of host cellular factors required for in uenza A virus replication
Buch in deiner Nähe kaufen
Beschreibung
The scope of this thesis was to further elucidate the function of the 287 primary hits identified in a primary genome-wide siRNA based-screen. These 287 human cellular factors had an effect on virus replication when knockdown studies were performed (Karlas et al., 2010). This study will investigate three major aspects:
The first part of this study focuses on the establishment and optimization of a validation assay to repeatedly screen the previously identified primary hits. Four different siRNAs will be utilized against each of the identified targets to minimize o-target effects. To investigate if the cellular factors are essential for a broad-range of IAVs, different IAV strains will be used for infection of the knockdown cells.
The activity of several hundred human cellular factors during virus replication are validated and confirmed in the validation screen. Several genes can also be referred to as a "druggable genome", which provides an exciting challenge to develop novel anti-viral agents against the influenza virus. This study focuses on the establishment and optimization of a small-molecules screening assay to identify anti-viral activity in a dose-dependent manner. More than 400 small-molecules shall be screened to target either specic human cell proteins or human kinases. This research will contribute to the identication of new targets for anti-viral agents.
Among the validated genes, p27 is one of the major hits and has furthermore been confirmed to have anti-viral effects in vivo (Karlas et al., 2010). p27 plays major roles in molecular and cellular mechanisms such as cell death, survival, and cell cycle. The focus of this study is to investigate the mechanism and function of p27 during the replication of the influenza virus. Human p27 knockdown and mouse p27 knockout cells will be utilized to investigate their phenotype after infection with IAV. In addition, the identification of a cellular or viral inducer of p27 expression will contribute to our understanding of the role of p27 during IAV replication.
Autor*in
Yu Jin Shin