Illuminating gene therapy

项目总结 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。弘扬严谨和 重复性，我们从单基因报告系统转向多重报告系统 可适用于广泛的基因编辑实施例的配置。具体来说，我们有 设计了一种由单体近红外荧光蛋白组成的适合于 深层组织测量，一种LacZ转基因基因，在整个器官和组织学中提供快速可见的染色 切片和实验室进化的红移荧光素酶，专为体内高度敏感而设计 测量。这种灵活的配置将使研究社区受益，提供多个读数 调查人员的选择。 所描述的鼠标被设计成检测包括胞嘧啶碱基的广泛的编辑实施例 编辑，腺嘌呤碱基编辑，Prime编辑，CasFx，也许还有其他。这种一体式设计将允许 研究人员将所需动物的数量降至最低，从而节省了时间和成本。它还促进了严谨和 重复性，因为使用相同动物的不同编辑技术可以更容易地量化 比较不同小组使用不同递送剂进行的研究和 CRISPR实施例。在撰写本文时，还没有这样的鼠标型号可用，因此这个项目将 为基因治疗领域提供了一种新颖独特的试剂。 我们已经组建了一个强大的团队来处理这个项目，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)。该项目还带来了一个 博士后研究员，在老鼠模型的基因治疗方面有专业知识(威尔逊·潘)，他开发了一种 为wtCas9基因编辑设计的鼠标传感器。不断增长的势头和不断增长的 有证据表明，基础编辑和主要编辑将成为临床竞赛的领跑者 治疗学。老鼠模型的发展将极大地加速我们将基因编辑带到 帮助确保更安全、更有效的治疗。

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.