Engineered Regulated RNA Localization and Transport in Biological Systems

描述(由申请人提供) 摘要：在许多生物体中，迅速积累的证据支持特定的和功能上有意义的亚细胞mRNA的定位。转录本定位影响许多基本过程，包括细胞极性、迁移和神经元活动，并影响发育、记忆和学习等不同的生物过程。虽然人们的注意力集中在几个选定的成绩单的本地化上，但最近的全球分析表明，这种现象非常普遍。在许多情况下，不同的转录本被观察到具有不同的亚细胞空间定位模式。一些突出的转录本定位形成过程的例子，如果蝇的体轴极性和神经元的突触可塑性，表明转录本定位可能具有直接的功能意义，现在被认为是一种普遍观察到的现象。不幸的是，我们对实现特定转录定位模式所需的蛋白质因子的了解非常有限。此外，我们缺乏有选择地干扰转录的空间分布的策略，这种策略与理解相关的功能后果相兼容。这项提议通过引入一种方法来满足这些需求，该方法允许将给定的转录本调节靶向亚细胞位置，后者由正在评估的已知或推定的本地化因素驱动。在这种方法中，转录定位完全由实验控制，并取决于小分子或肽信号分别应用于目标细胞的高分辨率化学梯度和翻译后蛋白质定位修饰。这一广泛适用的方法有可能促进我们对转录本定位的细胞机制的基础知识，并在细胞和生物水平上探索相关的功能含义。 公共卫生相关性：亚细胞RNA定位成离散的、依赖于转录的模式现在被认为是包括人类在内的许多细胞类型和生物体中的一种普遍现象。在几个研究得很好的案例中，转录定位是建立适当的细胞功能所必需的，这一过程中的缺陷会导致人类的发育、认知和其他神经问题。这项拟议的研究将引入新的方法来理解和操纵RNA亚细胞定位的潜在机制，并有可能改善我们对与RNA定位缺陷相关的疾病过程的理解和治疗。

DESCRIPTION (Provided by the applicant) Abstract: Evidence is rapidly accumulating in support of specific and functionally significant sub- cellular mRNA localization in many organisms. Transcript localization shapes many fundamental processes including cell polarity, migration and neuronal activity, and impacts diverse biological processes such as development, memory and learning. Although attention has been focused on localization of a few selected transcripts, recent global analyses indicate that the phenomenon is extremely common. In many cases, different transcripts are observed to have distinct sub-cellular spatial localization patterns. Several prominent examples of specific transcript localization shaping processes such as body axis polarity in Drosophila and synaptic plasticity in neurons suggest the potential for there being direct functional significance of transcript localization, now recognized to be a commonly observed phenomenon. Unfortunately, we have very limited understanding of the protein factors required for achieving specific transcript localization patterns. Moreover, we lack strategies for selectively perturbing transcript spatial distribution in a manner compatible with understanding the associated functional consequences. This proposal addresses these needs by introducing a method permitting regulated targeting of a given transcript to a sub-cellular location, the latter being driven by known or putative localization factors under evaluation. In this approach, transcript localization is entirely experimentally controlled, and is conditional upon either small molecule or peptide signals applied to target cells using high-resolution chemical gradients and post-translational protein localizing modifications, respectively. This broadly applicable method has the potential to advance our basic knowledge of the cellular mechanisms underlying transcript localization, and to probe the associated functional implications at both the cellular and organism levels. Public Health Relevance: Sub-cellular RNA localization into discrete, transcript-dependent patterns is now recognized to be a widespread phenomenon in many cell types and organisms, including humans. In several well-studied cases, transcript localization is required for establishing proper cellular function, and defects in this process contribute to developmental, cognitive and other neurological problems in humans. The proposed research will introduce new methods for understanding and manipulating the mechanisms underlying RNA sub-cellular localization, and has the potential to improve both our understanding and treatment of disease processes associated with RNA localization defects.