RNAi Mediated Gene Disruption in Caenorhabditis Elegans

申请以响应NIA先导研究资助计划的研究目标21“遗传学、行为、社会因素和老龄化”。虽然导致衰老的机制最近已成为一项密集的研究努力的主题，但衰老的多因素性质使人们很难理清导致衰老的因素。识别衰老原因机制的一个有效方法是使用深入分析的模式生物，如线虫秀丽线虫，研究延长寿命的基因突变或操作。一种新的识别线虫寿命基因的方法是使用RNA介导的推断(RNAi)。RNAi通过使用双链RNA来沉默基因表达，双链RNA针对特定的mRNAs进行降解。RNAi已被证明是研究线虫衰老的有价值的工具，并已被用于识别数百个影响线虫寿命的基因。然而，为了提供对衰老过程的真实洞察，对线虫和衰老的研究还必须分析其他生理功能指标，如代谢率，以及按时间顺序排列的寿命。在考虑延长线虫等外温(变温)生物体寿命的因素时，生理过程的测量尤其重要，因为降低外温生物代谢率的因素也往往会延长它们的寿命。这项研究将使用RNAi来操纵线虫的基因功能，并将这些操纵与这些动物的代谢率测量结合起来。最终，这种分析最有趣的结果之一将是揭示是否有任何经长寿命RNAi治疗的蠕虫也保持了正常的代谢率。这种蠕虫可以识别影响代谢率下游过程的候选基因，并为衰老的原因和减弱衰老的机制提供有价值的见解。

Application in response to research objective 21 "Genetics, Behavior, Social Factors and Aging" of the NIA pilot research grant program. While the mechanisms that cause aging have recently become the subject of an intensive research effort, the multi-factorial nature of aging makes it difficult to disentangle the factors responsible for senescence. A powerful method towards identifying the causal mechanisms of aging is to study genetic mutations or manipulations that extend longevity using an intensively analyzed model organism, such as the nematode Caenorhabditis elegans. A new method for identifying genes affecting longevity in C. elegans is the use of RNA-mediated inference (RNAi). RNAi silences gene expression through the use of double-stranded RNA that target specific mRNAs for degradation. RNAi has proven itself to be a valuable tool to study aging in C. elegans and has been used to identify hundreds of genes that influence longevity in C. elegans. However, to provide true insight into the aging process, studies of C. elegans and aging must also assay other measures of physiological function such as metabolic rates, in addition to chronological life span. Measures of physiological processes are especially important when considering factors that increase longevity in an ectothermic (poikilothermic) organism such as C. elegans because factors that reduce the metabolic rate of ectotherms also tend to increase their longevity. This research will use RNAi to manipulate gene function in C. elegans and to combine these manipulations with metabolic rate measurements on these animals. Ultimately one of the most interesting outcomes of such an analysis will be to reveal whether any of the longlived RNAi treated worms also retain normal metabolic rates. Such worms could identify candidate genes affecting processes that act "downstream" from the metabolic rate and provide valuable insights into the causes of aging and mechanisms by which it can be attenuated.