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

信使RNA(MRNAs)编码制造蛋白质的指令，蛋白质构成细胞功能的基本机制。最近的技术进步使可传递给人类的治疗性mRNAs的开发成为可能，这在广泛使用的SARS-CoV-2疫苗中尤为明显。这些疗法的成功表明，新一代的信使核糖核酸药物具有超越疫苗应用的潜力，例如抗癌治疗和遗传疾病的治疗。要实现这些目标，将需要设计优化蛋白质表达的mRNAs，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个RNA的合成库的翻译功能，以发现改变蛋白质生产的时间和数量的特征。对这些基因表达基本规则的洞察将是设计新类别的信使核糖核酸疗法的重要基石，以解决更广泛的人类疾病，从而推进RNA生物技术。该项目还将为博士后学者提供培训机会，并让初中生参与RNA生物学。从RNA合成的蛋白质数量可以跨越两个数量级以上，不同细胞类型之间存在差异，并随着细胞信号的变化而快速变化。决定这一表达范围的mRNAs的功能在很大程度上仍不清楚。可能的可能性包括由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。该项目将使用最近开发的大规模平行报告分析来系统地识别数千个人类和病毒5‘UTRs的特征，这些特征决定了蛋白质的产量和时间。目标1将检查不同类别的核苷酸修饰的翻译功能，并确定识别它们的分子机制。AIM 2将量化从环状RNA合成文库中产生的蛋白质，以确定能够访问非规范翻译启动机制的RNA元件。目标3将确定决定细胞类型特异性表达的mRNAs的特征以及建立这种特异性的机制。这些工作将提供对mRNA编码的翻译决定因素的全面了解，提供对基因表达的基本决定因素的洞察，并指导新的mRNA疗法的设计。该项目得到了生物科学局的遗传机制计划/分子和细胞生物科学部以及数学和物理科学局的化学部的支持。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.