MFB: Massively parallel identification of translation regulatory sequences in human and viral mRNAs

信使rna （mrna）编码制造蛋白质的指令，构成细胞功能的基本机制。最近的技术进步使得能够开发出可用于人体的治疗性mrna，特别是在广泛使用的SARS-CoV-2疫苗中。这些疗法的成功表明，新一代mRNA药物在疫苗之外的应用潜力，如抗癌疗法和遗传疾病的治疗。实现这些目标将需要设计优化蛋白质表达的mrna，并且可以针对特定的组织和细胞环境进行定制。该项目的目标是利用一种策略来量化数千种rna合成文库的翻译功能，以发现改变蛋白质生产时间和数量的特征。深入了解基因表达的这些基本规则，将成为开发新型mRNA治疗药物的重要基石，以解决更广泛的人类疾病，从而推动RNA生物技术的发展。该项目还将为博士后学者提供培训机会，并吸引初高中学生参与RNA生物学。由mRNA合成的蛋白质的数量可以跨越两个数量级以上，在不同的细胞类型中变化，并且在响应细胞信号时迅速变化。决定这种表达范围的mrna的特征在很大程度上仍然未知。可能性包括RNA结合蛋白识别的序列基序、结构RNA元件和影响翻译过程的核苷酸修饰。该项目将采用最近开发的大规模平行报告基因试验，系统地识别数千种人类和病毒5 ' utr的特征，这些特征决定了蛋白质生产的数量和时间。目的1将研究不同种类的核苷酸修饰的翻译功能，并确定识别它们的分子机制。目的2将从环状RNA合成文库中量化蛋白质的产生，以鉴定能够进入非规范翻译起始机制的RNA元件。目的3将确定mrna决定细胞类型特异性表达的特征和建立这种特异性的机制。这些努力将提供对mRNA编码的翻译决定因素的全面理解，为基因表达的基本决定因素提供见解，并指导新的mRNA治疗方法的设计。该项目得到了遗传机制项目/生物科学理事会分子和细胞生物科学部以及数学和物理科学理事会化学部的支持。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Messenger RNAs (mRNAs) encode instructions for making proteins, which constitute the fundamental machinery for cellular function. Recent technological advancements have enabled the development of therapeutic mRNAs that can be delivered to humans, notably evident in widely used vaccines for SARS-CoV-2. The success of these therapeutics suggests the potential of new generations of mRNA medicines with applications beyond vaccines, such as anti- cancer therapies and treatment for genetic disorders. Realizing these goals will require the design of mRNAs that optimize protein expression and can be customized for specific tissues and cellular environments. The goal of this project is to leverage a strategy for quantifying the translation functions of synthetic libraries of thousands of RNAs to discover features that modify the timing and quantity of protein production. Insights into these fundamental rules for gene expression will be important building blocks for engineering new classes of mRNA therapeutics to address a broader spectrum of human disease, thus advancing RNA biotechnology. The project will also provide training opportunities for postdoctoral scholars and engage middle and high school students in RNA biology.The quantity of protein synthesized from an mRNA can span more than two orders of magnitude, vary across cell types, and rapidly change in response to cellular signals. The features of mRNAs that determine this range of expression remain largely unknown. Possibilities include sequence motifs recognized by RNA-binding proteins, structured RNA elements and nucleotide modifications that affect the translation process. This project will employ a recently developed massively parallel reporter assay to systematically identify features of thousands of human and viral 5′ UTRs that determine the amount and timing of protein production. Aim 1 will examine the translation functions of diverse classes of nucleotide modifications and identify the molecular mechanisms that recognize them. Aim 2 will quantify protein production from a synthetic library of circular RNAs to identify RNA elements capable of accessing mechanisms for non-canonical translation initiation. Aim 3 will identify features of mRNAs that determine cell-type specific expression and the mechanisms that establish this specificity. These efforts will provide a comprehensive understanding of mRNA- encoded determinants of translation, providing insights into basic determinants of gene expression and guiding the design of new mRNA therapeutics.This project is supported by the Genetic Mechanisms program/Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences and by the Division of Chemistry in the Directorate for Mathematical and Physical Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.