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

MFB：大规模并行鉴定人类和病毒 mRNA 中的翻译调控序列

• 批准号: 2330451
• 负责人: Carson Thoreen
• 金额: $ 144万
• 依托单位: Yale University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2330451&HistoricalAwards=false
• 关键词: MFB; Massively; parallel; identification; translation

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.

Messenger RNA（mRNA）编码制造蛋白质的说明，这构成了细胞功能的基本机制。最近的技术进步使可以将治疗性mRNA的发展开发给人类，这尤其是在SARS-COV-2的广泛使用疫苗中的证据。这些治疗剂的成功表明，新一代的mRNA药物的潜力以及疫苗以外的应用，例如抗癌疗法和实现这些目标的治疗，将需要设计优化蛋白质表达的mRNA，并可以针对特定的组织和细胞环境进行定制。该项目的目的是利用量化成千上万RNA的合成库的翻译功能的策略，以发现修改蛋白质产生的时间和数量的特征。对这些基因表达的这些基本规则的见解将是工程新的mRNA疗法的重要组成部分，以解决更广泛的人类疾病，从而提高了RNA生物技术。该项目还将为博士后学者提供培训机会，并吸引中学生参与RNA生物学。从mRNA合成的蛋白质数量可以跨越两个以上的数量级，各种细胞类型各不相同，并且会因细胞信号而迅速变化。确定这种表达范围的mRNA的特征在很大程度上仍然未知。可能性包括通过RNA结合蛋白识别的序列基序，影响翻译过程的结构化RNA元素和核修饰。该项目将采用最近开发的大量平行记者测定法，以系统地识别数千个人和病毒5'UTR的特征，这些特征决定了蛋白质产生的数量和时机。 AIM 1将检查核苷酸修饰的潜水员类别的翻译功能，并确定识别它们的分子机制。 AIM 2将从圆形RNA的合成库中量化蛋白质的产生，以识别能够访问非典型翻译起始机制的RNA元素。 AIM 3将确定确定细胞类型特异性表达的mRNA特征和建立这种特异性的机制。这些努力将为MRNA的确定性提供全面的理解，从而提供了基本确定基因表达的见解，并指导新的mRNA疗法的设计。该项目得到了遗传机制计划/分子和细胞生物科学在生物科学和物理派系中的分子和细胞裁决的分子和细胞生物科学的划分的支持，并由NESForate and Science and Nations and Issions and Issions and Issive and Issive and Nations. Matheration. Mathistory Sciity and. Mathistory Sciity。使用基金会的知识分子优点和更广泛的审查标准，通过评估被认为值得支持。