Developing Next Generation Genetics for Understanding Hematopoietic Stem Cell Biology

项目总结： 基于Cre重组酶和Cas9核酸酶的小鼠遗传工具已被应用于细胞/组织特异性 基因敲除和细胞谱系追踪。这些工具能够控制基因表达和基因组 工程学，揭示许多系统中的详细生物学。然而，缺乏精确的时空 控件阻碍了这些工具的更广泛应用。在这个提案中，我们将开发一套新鼠标 解决这一问题的基因工具。然后，我们建议应用这一制度来调查建立 体内成人骨髓造血干细胞库。因为造血干细胞可以自我更新和分化 在整个生命过程中，它们是挽救生命的骨髓移植的基础 诊所。因此，了解HSC生物学具有重要的翻译意义。成立了 成人骨髓HSC库对终生造血是必不可少的。然而，成人的骨髓是如何 HSC池是否已建立尚未得到解决。缺乏适当的遗传工具来研究造血干细胞 体内克隆水平一直是一个主要障碍。我们的新基因工具将允许在克隆水平上追踪造血干细胞 在活体内。如果成功，结果将通过直接解决成人骨骼如何 建立骨髓间充质干细胞池。此外，我们提出的新技术将广泛适用于各种 研究领域超越了造血，以解决根本问题。

Project Summary: Mouse genetic tools based on Cre recombinase and Cas9 nuclease have been applied for cell/tissue-specific gene knockout and cell lineage tracing. These tools enable control of gene expression and genome engineering to uncover detailed biology in a number of systems. However, the lack of precise spatiotemporal control hinders the broader applications of these tools. In this proposal, we will develop a new set of mouse genetic tools to address this issue. Then we propose to apply this system to investigate the establishment of the adult bone marrow hematopoietic stem cell (HSC) pool in vivo. Because HSCs self-renew and differentiate into all blood cell lineages throughout life, they are the basis for life-saving bone marrow transplantation in clinics. Therefore, understanding HSC biology has important translational implications. The establishment of the adult bone marrow HSC pool is essential for life-long hematopoiesis. However, how the adult bone marrow HSC pool is established has not been addressed. The lack of appropriate genetic tools to study HSCs at the clonal level in vivo has been a major hurdle. Our novel genetic tools will allow tracing HSCs at the clonal level in vivo. If successful, the results will significantly advance the field by directly addressing how the adult bone marrow HSC pool is established. Also, our proposed novel technologies will be widely applicable to various research fields beyond hematopoiesis to address fundamental questions.