Genome-wide RNA interference screen for the identification of crucial host cell factors in Chlamydia trachomatis infection
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Chlamydia trachomatis is a bacterial pathogen with major impact on human health; it is responsible for diseases such as blinding trachoma and the most common sexually transmitted diseases that can cause infertility if not treated successfully. Despite the wide range of clinical manifestations, little is known about the mechanisms underlying the infection process, which depends on extensive interactions with the host cell allowing Chlamydia to acquire critical nutrients and to avoid host anti-microbial defenses. Due to fact that Chlamydia is not yet amenable to routine manipulation with standard molecular genetic tools that have been central to most studies in bacterial pathogenesis, the functional analysis of Chlamydia infection is focused on the role of the host cell. Increasing infection rates and emerging antibiotic resistance in Chlamydia further substantiate the urgency of a better understanding of the molecular bases of the infection processes. Aim of the present study was to develop a strategy to investigate the role of host cell factors in Chlamydia infection on a genome-wide scale using RNA interference technology. In this line, a cell-based fluorescent read-out system had to be established to quantify the production of Chlamydia infectious progeny (infectivity) upon host cell knockdown conditions. The assay was to be transferred to an automatic robotic platform and improved for its robustness and reliability in order to be able to perform a high-throughput siRNA screen on a genome-wide scale. The identified host cell factors with essential functions in Chlamydia infection were to be validated and categorized regarding their location, function, and their role in molecular processes.
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Genome-wide RNA interference screen for the identification of crucial host cell factors in Chlamydia trachomatis infection, Erik González
- Sprache
- Erscheinungsdatum
- 2013
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- Titel
- Genome-wide RNA interference screen for the identification of crucial host cell factors in Chlamydia trachomatis infection
- Sprache
- Englisch
- Autor*innen
- Erik González
- Verlag
- mbv
- Erscheinungsdatum
- 2013
- ISBN10
- 3863874145
- ISBN13
- 9783863874148
- Reihe
- Biology
- Kategorie
- Medizin & Gesundheit
- Beschreibung
- Chlamydia trachomatis is a bacterial pathogen with major impact on human health; it is responsible for diseases such as blinding trachoma and the most common sexually transmitted diseases that can cause infertility if not treated successfully. Despite the wide range of clinical manifestations, little is known about the mechanisms underlying the infection process, which depends on extensive interactions with the host cell allowing Chlamydia to acquire critical nutrients and to avoid host anti-microbial defenses. Due to fact that Chlamydia is not yet amenable to routine manipulation with standard molecular genetic tools that have been central to most studies in bacterial pathogenesis, the functional analysis of Chlamydia infection is focused on the role of the host cell. Increasing infection rates and emerging antibiotic resistance in Chlamydia further substantiate the urgency of a better understanding of the molecular bases of the infection processes. Aim of the present study was to develop a strategy to investigate the role of host cell factors in Chlamydia infection on a genome-wide scale using RNA interference technology. In this line, a cell-based fluorescent read-out system had to be established to quantify the production of Chlamydia infectious progeny (infectivity) upon host cell knockdown conditions. The assay was to be transferred to an automatic robotic platform and improved for its robustness and reliability in order to be able to perform a high-throughput siRNA screen on a genome-wide scale. The identified host cell factors with essential functions in Chlamydia infection were to be validated and categorized regarding their location, function, and their role in molecular processes.