Illuminating gene therapy

照亮基因疗法

基本信息

批准号：
10573764
负责人：
MICHAEL T MCMANUS
金额：
$ 20.19万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10573764
关键词：
Acceleration Adenine Animal Model Animals CRISPR/Cas technology Cell model Cells Clinic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Communities Cytosine DNA cassette Data Data Reporting Development ES Cell Line Engineering Ensure Fluorescence Genes Goals Histology Laboratories LacZ Genes Liver Luciferases Measurement Modality Mus Nature Organ Outcome Patients Positioning Attribute Postdoctoral Fellow Proteins Race Reagent Reporter Reporter Genes Reproducibility Research Research Personnel Safety Solid Neoplasm Stains System Technology Testing Therapeutic Time Tissues Transgenes Transgenic Organisms Validation Visual Work Writing base editing base editor clinical application cost design ds-DNA efficacious treatment embryonic stem cell experimental study flexibility gene therapy human disease in vivo monomer mouse development mouse genetics mouse model novel prime editing prime editor redshift research study sensor standard care

项目摘要

PROJECT SUMMARY In this project, we will develop a new mouse model for quantitating gene therapy. To promote rigor and reproducibility, we move away from single gene reporter systems and transition to multiplexed reporter configurations that are amenable for a broad range of gene editing embodiments. Specifically, we have designed an expression cassette composed of a monomeric near-infrared fluorescence protein suitable for deep tissue measurements, a LacZ transgene that provides a rapid visible stain in whole organs and histology sections, and laboratory evolved red-shifted luciferase that is designed for highly sensitive in vivo measurements. This flexible configuration will benefit the research community, offering multiple readout options for investigators. The described mouse is engineered to detect a broad range of editing embodiments including Cytosine Base Editors, Adenine Base Editors, Prime-Editors, CasFx, and perhaps others. This ‘all-in-one’ design will allow researchers to minimize the number of animals needed, thus saving time and cost. It also promotes rigor and reproducibility since different editing technologies that use the same animal can be more readily quantitatively compared between research studies performed by different groups, using different delivery agents and CRISPR embodiments. As of this writing, there are no such mouse models available, hence this project will provide a novel and unique reagent for the gene therapy field. We have assembled a strong team to tackle this project, guided by experts in base editors and prime editors (David Liu and Jennifer Doudna) and mouse genetics (Michael McManus). The project also brings on board a postdoc co-investigator with expertise in gene therapy in mouse models (Wilson Poon) who has developed a mouse sensor designed for wtCas9 gene editing. Growing momentum and an ever-increasing body of evidence indicates that base editors and prime editors will become front-runners in the race to clinical therapeutics. The development of mouse models will greatly accelerate our goal to bring gene editing to the clinic, helping to ensure a safer and more efficacious treatment.

项目摘要在这个项目中，我们将开发一种新的小鼠模型来定量基因治疗。促进严格和可重复性，我们从单个基因报告基因系统转移到多路复用报告基因适用于广泛基因编辑实施方案的构型。具体来说，我们有设计了一个由单体近红外荧光蛋白组成的表达盒深组织测量值，一种LACZ变换，可在整个器官和组织学中迅速可见染色部分和实验室进化的红移荧光素酶，专为体内高度敏感测量。这种灵活的配置将使研究社区受益，并提供多个读数调查人员的选项。所描述的小鼠经过设计以检测包括胞嘧啶碱基在内的广泛的编辑实施方案编辑，腺嘌呤基础编辑，主要编辑，CASFX以及其他人。这种“多合一”设计将允许研究人员可以最大程度地减少所需的动物数量，从而节省时间和成本。它还促进了严格和可重复性，因为使用同一动物的不同编辑技术可以更容易定量比较不同群体进行的研究，使用不同的递送剂和 CRISPR实施例。在撰写本文时，没有这样的鼠标模型，因此这个项目将为基因治疗领域提供一种新颖而独特的试剂。我们已经组建了一个强大的团队来解决该项目，在基础编辑和主要编辑的专家的指导下（David Liu和Jennifer Doudna）和老鼠遗传学（Michael McManus）。该项目还带来了A DOSDOC共同研究器具有基因疗法专业知识的小鼠模型（Wilson Poon），后者已经开发了为WTCAS9基因编辑设计的小鼠传感器。增长的动力和不断增长的身体证据表明基本编辑和主要编辑将成为临床比赛的领先者疗法。鼠标模型的开发将极大地加速我们的目标，以将基因编辑带入诊所，有助于确保更安全，更有效的治疗方法。