Analysis of the C.albicansTranscriptome

描述(由申请人提供)：该项目将通过使用超高通量RNA测序，结合合理选择的不同实验条件，准确地确定人类真菌病原体白色念珠菌基因组中所有转录本的身份和位置。白念珠菌是一种日益重要的人类病原体，由于患者人数的增加和对现有抗真菌药物的耐药性增加，开发新的治疗方法至关重要。使用我们的方法可能会发现数百个新的转录本，其中许多可能是白色念珠菌或密切相关物种的特异性基因；这些新的病原体特异性基因可能被证明是很好的治疗靶点。此外，我们将通过RT-PCR验证所有新的转录本，并确定哪些新的转录本可能编码蛋白质。最后，我们将对新的转录本进行初步的功能表征，通过使用微阵列来确定它们在各种使用的实验条件下的相对丰度，并通过确定它们与哪些已知功能的基因共同调节。这一建议具有重要意义，既是因为它直接促进了我们对这种医学上重要的人类真菌病原体的生物学的理解，也是因为来自其他生物的类似研究强烈表明，拟议的实验将大大完善白色念珠菌基因组的主要注释。因此，我们获得了对白色念珠菌生物学的洞察，也使真菌基因组学领域更广泛地受益，长期目标是对人类患者治疗真菌感染的方式产生重大影响。随着白念珠菌基因组的染色体水平组装，高通量测序技术的存在，以及公共念珠菌基因组学数据库的存在，为成功完成这一项目并将数据传播到真菌发病机制研究社区奠定了基础，这将对未来的白念珠菌研究，并最终治疗念珠菌疾病产生重大影响。 公共卫生相关性：白色念珠菌是一种日益重要的人类病原体，开发新的治疗方法至关重要，特别是考虑到最近白色念珠菌临床分离株的耐药性上升。这项工作旨在识别白色念珠菌中的新基因；很可能会识别数百个新基因，其中许多可能被证明是很好的治疗靶点。因此，这项工作有可能在未来对人类健康产生巨大的积极影响。

DESCRIPTION (provided by applicant): This project will define precisely the identity and positions of all transcripts within the genome of the human fungal pathogen Candida albicans, by using ultra high throughput RNA sequencing, in conjunction with a rationally chosen diverse set of experimental conditions. C. albicans is an increasingly important human pathogen, and, due to an expanding patient population and to increasing resistance to existing antifungal drugs, it is vital that new treatments are developed. It is likely that hundreds of new transcripts will be identified using our approach, and many of these may be specific to C. albicans or closely related species; these novel pathogen-specific genes may prove to be good therapeutic targets. In addition, we will validate all new transcripts by RT-PCR, and identify which of the new transcripts are likely to encode proteins. Finally, we will perform preliminary functional characterization of the novel transcripts, by using microarrays to determine their relative abundance under the various used experimental conditions, and by identifying with which genes of known function they are co-regulated. This proposal is significant both because it directly advances our understanding of the biology of this medically important human fungal pathogen, and because similar studies from other organisms strongly suggest that the proposed experiments will substantially refine the primary annotation of the C. albicans genome. We thus gain insight into C. albicans biology and also benefit the field of fungal genomics more widely, with the long-term goal of having a significant impact into the way in which fungal infections are treated in human patients. With a chromosome-level assembly of the C. albicans genome, the existence of high throughput sequencing technologies, and a public Candida genomics database, the groundwork is in place for both successful completion of this project and dissemination of the data to the fungal pathogenesis research community to significantly impact future C. albicans research, and, ultimately the treatment of Candida disease. PUBLIC HEALTH RELEVANCE: Candida albicans is an increasingly important human pathogen, and the development of new treatments is vital, especially given the recent rise in drug resistance among clinical isolates of C. albicans. This work seeks to identify novel genes within C. albicans; it is likely that hundreds of new genes will be identified, and of those, many may prove to be good therapeutic targets. Thus, this work has the potential for a large positive future impact on human health.