Therapeutic strategies for microsatellite expansion diseases using RNA-targeting CRISPR/Cas

使用 RNA 靶向 CRISPR/Cas 治疗微卫星扩增疾病的策略

基本信息

批准号：
10171924
负责人：
MAURICE SCOTT SWANSON
金额：
$ 59.05万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-22 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10171924
关键词：
3&apos Untranslated Regions Adopted Adult Affect Alternative Splicing Animal Model Autopsy BCAR1 gene Binding Biological Markers Biopsy C9ORF72 Cell Extracts Cell model Cells Characteristics Chemicals Cleaved cell Clinical Clustered Regularly Interspaced Short Palindromic Repeats DNA Data Defect Dependovirus Development Disease Endoribonucleases Event Foundations Genes Genetic Transcription Goals Guide RNA Health Human Huntington Disease Image In Vitro Inherited Introns Knock-in Mouse Lead Link Mammalian Cell Measures Mediating Messenger RNA Microsatellite Instability Microsatellite Repeats Modeling Modification Movement Mus Muscle Muscle Fibers Muscular Dystrophies Myoblasts Myotonic Dystrophy Neuromuscular Diseases Nuclear RNA Oligonucleotides Pathogenesis Patients Polyadenylation Proteins RNA RNA Binding RNA Degradation RNA Processing RNA-Binding Proteins Safety Serotyping Specificity System Technology Testing Therapeutic Time Tissues Toxic effect Transduction Gene Transgenes Treatment Efficacy Viral Genome adeno-associated viral vector base biomarker panel efficacy evaluation efficacy study gene therapy genome editing human disease human subject immunogenicity in vivo mouse model novel participant safety pre-clinical prognostic programs recruit safety study stress granule success therapeutic development therapeutic evaluation therapeutic target therapy development transcriptome sequencing

项目摘要

Project Summary The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas system has become widely adopted for DNA recognition, enabling applications such as genome editing and recruiting effector proteins to DNA to affect transcription or enable imaging. Recently in Nelles et al, Cell, 2016, we showed that with modifications, catalytically dead Cas9 with a modified oligonucleotide containing the PAM recognition sequence is able to bind specific mRNAs in living mammalian cells and is able to track their movement, opening up the potential for many RNA applications of Cas proteins. In this proposal, we aim at developing a adeno-associated virus (AAV)-based therapeutic strategy for myotonic dystrophy, a microsatellite expansion disease characterized by expanded CTG repeats, using RNA-targeting CRISPR/Cas9 (RCas9). We will perform in vivo safety and efficacy studies of our AAV-based therapy and evaluate the hypothesis that alternative RNA processing biomarkers are reliable for measuring treatment efficacy of RCas9 in myotonic dystrophy. If successful, we will have taken a significant step forward in developing a treatment for a class of microsatellite expansion diseases.!

项目概要成簇规则间隔短回文重复序列 (CRISPR)/Cas 系统已得到广泛应用用于 DNA 识别，使基因组编辑和招募效应蛋白等应用成为可能 DNA 影响转录或实现成像。最近，在 Nelles 等人的《Cell》，2016 年中，我们证明了修饰，带有包含 PAM 识别的修饰寡核苷酸的催化死亡 Cas9 序列能够结合活体哺乳动物细胞中的特定 mRNA，并能够追踪它们的运动，为 Cas 蛋白的许多 RNA 应用开辟了潜力。在本提案中，我们的目标是开发一个基于腺相关病毒（AAV）的强直性肌营养不良治疗策略，微卫星扩展使用 RNA 靶向 CRISPR/Cas9 (RCas9) 来扩展 CTG 重复序列为特征的疾病。我们将对我们基于 AAV 的疗法进行体内安全性和有效性研究，并评估以下假设：替代 RNA 加工生物标志物可可靠地测量 RCas9 在肌强直中的治疗效果营养不良。如果成功的话，我们将在开发针对一类疾病的治疗方法方面迈出重要一步。微卫星扩张疾病。！