Advancing stem cell technologies using engineered zinc finger proteins

使用工程锌指蛋白推进干细胞技术

基本信息

批准号：
8143426
负责人：
J. KEITH JOUNG
金额：
$ 87.62万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-30 至 2012-07-31
项目状态：
已结题

项目摘要

DESCRIPTION Abstract: Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), with their abilities to replicate indefinitely and maintain pluripotency, offer tremendous opportunities for the development of novel regenerative medicine-based therapies. iPSC technology, in particular, may provide the means to develop personalized therapeutic strategies in which a patient's own cells might be modified and used to treat their disease without the need for immunosuppression. Patient-derived ESCs and iPSCs will also provide critically important reagents for modeling and studying the biology of various diseases. Despite tremendous recent progress, many significant challenges remain to be addressed before ESCs and iPSCs can be used routinely for therapeutics. For example, methods for generating iPSCs are currently inefficient with considerable room for improvement. In addition, homologous recombination-mediated gene targeting for creating specific mutations works with low efficiency and is difficult to perform, particularly in human ESCs and iPSCs. Lastly, methods for robustly differentiating ESCs and iPSCs into cell types of interest are typically inefficient and yield cells that fail to exhibit all desired characteristics. The goal of this proposal is to employ engineered zinc finger technology to address these major challenges impeding research and therapeutic applications of ESCs and iPSCs. Specifically, this proposal will utilize the PI's expertise in engineered zinc finger technology to address three important challenges for stem cell biology and research: (1) developing more efficient methods for iPSC generation, (2) enabling high efficiency ZFN-induced gene targeting in human ESCs and iPSCs, and (3) creating more efficient methods for differentiating stem cells into desired cell types of interest.

描述抽象的：胚胎干细胞（ESC）和诱导多能干细胞（IPSC），其能力无限期地复制并保持多能性，为开发新的基于再生医学的疗法提供了巨大的机会。尤其是IPSC技术，可以提供开发个性化治疗策略的手段，其中可以修改患者自己的细胞并用于治疗其疾病而无需免疫抑制。患者来源的ESC和IPSC还将为建模和研究各种疾病的生物学建模和研究至关重要的试剂。尽管最近取得了巨大进展，但在ESC和IPSC可以定期用于治疗之前，仍有许多重大挑战要解决。例如，生成IPSC的方法当前效率低下，有相当大的改进空间。此外，与创建特定突变的同源重组介导的基因靶向效率低，并且难以执行，尤其是在人类ESC和IPSC中。最后，将ESC和IPSC鲁棒分化为感兴趣的细胞类型的方法通常是低效的，而产量细胞无法表现出所有所需的特征。该提案的目的是采用工程化的锌指技术来应对妨碍ESC和IPSCS的研究和治疗应用所面临的主要挑战。具体而言，该建议将利用PI在工程锌指技术方面的专业知识来应对干细胞生物学和研究的三个重要挑战：（1）为IPSC生成开发更有效的方法，（2）能够高效ZFN诱导的基因靶向人类和IPSC中的ESC和IPSC，以及（3）对较高的细胞进行兴趣分类的细胞，从而使型细胞更加有效。