Engineered promoters for finely tuned gene expression in Chlamydia

摘要 沙眼衣原体是一个重要的健康问题，在世界范围内。大幅 负担的存在是由于流行和综合健康和社会经济影响， 急性和慢性疾病。衣原体是专性细胞内寄生虫， 复杂的发育周期，这种生物学在历史上阻碍了快速发展， 了解发病机制。最近，用外源DNA转化衣原体的能力 极大地促进了对支持这些疾病的分子机制的理解， 衣原体感染成功。例如，衣原体基因的异位表达， 提供了对细胞内定位和功能的深入了解。随着研究越来越多， 然而，目前复杂的启动子系统常常被证明不足以解决 生理相关的生物学。进展特别受到限制的选择，在严格- 受调控的诱导型启动子。我们建议通过开发试剂来解决缺陷， 更精细的基因表达。我们将利用合成生物学的进步 和合理的设计来设计一套严格调节和可调的启动子。初步 本文提供了显示该方法的功效的数据。我们建议验证和 进一步发展这些技术。在这些研究结束时，我们将建立新的 这些方法将使整个衣原体研究界受益，并促进 异位基因表达研究衣原体感染生物学。

ABSTRACT Chlamydia trachomatis represents a significant health concern world-wide. A substantial burden exists due to the prevalence and the combined health and socioeconomic impact of acute and chronic disease. Chlamydiae are obligate intracellular parasites that undergo a complex developmental cycle, and this biology has historically impeded rapid progress in understanding pathogenesis. Recently, the ability to transform Chlamydia with exogenous DNA has greatly facilitated advances in understanding the molecular mechanisms underpinning the success of chlamydial infection. For example, ectopic expression of chlamydial genes has provided insight into intracellular localization and function. As studies become more sophisticated, however, current promoter systems are often proving inadequate to address physiologically relevant biology. Progress is particularly hindered by limited choices in tightly- regulated, inducible promoters. We propose to address deficiencies by developing reagents for more finely tuned gene expression in Chlamydia. We will exploit advances in synthetic biology and rational design to engineer a suite of tightly regulated and tunable promoters. Preliminary data showing the efficacy of the approach are presented herein. We propose to validate and further develop these technologies. At the end of these studies, we will have established new approaches that will benefit the entire Chlamydia research community and advance the utility of ectopic gene expression to study chlamydial infection biology.