Molecular tools for targeting RNA

用于靶向 RNA 的分子工具

基本信息

批准号：
10393058
负责人：
Patrick Hsu
金额：
$ 17.97万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2022-10-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10393058
关键词：
Adopted Alternative Splicing Amino Acids Binding Biochemical Bioinformatics Biological Models Biomedical Research CRISPR/Cas technology Catalogs Cell physiology Cells Clustered Regularly Interspaced Short Palindromic Repeats Code Communities Complement DNA DNA Binding Domain Deoxyribonucleases Dependovirus Development Diagnostic Disease Elements Engineering Enzymes Foundations Frontotemporal Dementia Future Gene Expression Genes Genetic Diseases Genetic Transcription Genetic Variation Genetic study Genome Genome engineering Genomics Goals Guide RNA Human Image Individual Knock-in Light Mainstreaming Mammalian Cell Maps Modeling Molecular Neurons Nucleic Acids Nucleotides Pancreatic ribonuclease Patients Phenotype Protein Engineering Protein Isoforms RNA RNA Binding RNA Interference RNA Splicing Regulation Research Ribonucleases Scientist Site Small RNA Structure System Tau isoform ratio Technology Therapeutic Transcript Untranslated RNA Variant Viral Viral Vector Work adeno-associated viral vector base delivery vehicle epigenome editing expression vector flexibility genome editing genomic locus induced pluripotent stem cell interest knock-down knockout gene next generation sequencing novel nuclease optogenetics programs public health relevance spatiotemporal stem cell differentiation therapeutic development tool transcriptome transcriptome sequencing

项目摘要

Abstract Due to rapid advances in RNA sequencing, scientists now have the ability to easily map transcriptome changes in cellular function and disease to complement a robust catalog of functional genome elements and variants. Genome editing tools, in particular the CRISPR-Cas9 platform, allow for rapid control and manipulation of genome sequences for genetic study. To extend these tools to the direct and facile perturbation of RNA, the proposed work aims to develop a broad suite of transcriptome engineering tools based on diverse RNA-targeting CRISPR systems to complement RNAi. Analogous to dCas9 for DNA, programmable RNA binding will enable modular modes of function including and beyond RNA knockdown, such as splice isoform engineering, by leveraging a combination of bioinformatic, biochemical, and protein engineering approaches to demonstrate and optimize the utility of the system. Using patient-derived induced pluripotent stem cells differentiated into neurons as a model system, RNA mis-splicing diseases will be targeted as a proof-of-concept to modulate alternative splicing. This proposal expects to enable a robust, flexible platform to interrogate gene expression, study transcript dynamics, and dissect the function of coding and noncoding transcripts.

抽象的由于 RNA 测序技术的快速进步，科学家现在能够轻松绘制转录组图谱细胞功能和疾病的变化，以补充功能基因组元件的强大目录和变种。基因组编辑工具，特别是 CRISPR-Cas9 平台，可以快速控制和操纵基因组序列进行遗传研究。将这些工具扩展到直接和方便 RNA 的扰动，拟议的工作旨在开发一套广泛的转录组工程工具基于多种 RNA 靶向 CRISPR 系统来补充 RNAi。类似于 DNA 的 dCas9，可编程RNA结合将实现模块化功能模式，包括RNA敲除，例如通过利用生物信息学、生物化学和蛋白质的组合进行剪接异构体工程展示和优化系统实用性的工程方法。使用源自患者的诱导以多能干细胞分化为神经元为模型系统，RNA错误剪接疾病将旨在作为调节选择性剪接的概念验证。该提案期望能够建立一个强大的、灵活的平台来询问基因表达、研究转录动态并剖析编码功能和非编码转录本。