Developing Next Generation Genetics for Understanding Hematopoietic Stem Cell Biology

开发下一代遗传学以了解造血干细胞生物学

• 批准号: 10710163
• 负责人: Lei Ding
• 金额: --
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-27 至 2023-07-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10710163
• 关键词: Address; Adult; Bar Codes; Binding; Biological Process; Biology; Bioluminescence; Blood; Blood Cells; Bone Marrow; Bone Marrow Transplantation; CRISPR/Cas technology; Cell Lineage; Cells; Clinic; Clonal Hematopoietic Stem Cell; Codon Nucleotides; Communities; DNA Binding Domain; Development; Disease; Dissociation; Doxycycline; Enterobacteria phage P1 Cre recombinase; Fluorescence; Gene Expression; Genes; Genetic; Genetic Recombination; Genetic Transcription; Genome engineering; Goals; Guide RNA; HSF1; Hematopoiesis; Hematopoietic stem cells; In Vitro; Libraries; Life; Light; Lighting; Liver; Mammalian Cell; Mammals; Molecular; Mus; Names; Newborn Infant; Nuclear Localization Signal; Pathologic; Photoreceptors; Physiologic pulse; RNA; Reporter; Research; Resolution; Site; Sorting; Specificity; Stimulus; System; Tamoxifen; Technology; Testing; Tetracyclines; Tissues; Transcriptional Activation Domain; Work; base; experience; experimental study; gene repression; genetic manipulation; genetic technology; improved; in vivo; innovation; knock-down; knockout gene; mouse genetics; mutant; new technology; next generation; novel; nuclease; optogenetics; p65; self renewing cell; single-cell RNA sequencing; small molecule; spatiotemporal; stem cell biology; stem cell migration; synthetic biology; tool

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.

项目概要： 基于Cre重组酶和Cas9核酸酶的小鼠遗传工具已应用于细胞/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 工程以揭示许多系统中的详细生物学。但缺乏精确的时空 控制阻碍了这些工具的更广泛应用。在这个提案中，我们将开发一套新的鼠标 遗传工具可以解决这个问题。然后我们建议应用这个系统来调查建立 体内成人骨髓造血干细胞（HSC）池。因为 HSC 会自我更新和分化 它们进入整个生命周期的所有血细胞谱系，是挽救生​​命的骨髓移植的基础 诊所。因此，了解 HSC 生物学具有重要的转化意义。成立 成人骨髓 HSC 池对于终生造血至关重要。然而，成人骨髓如何 HSC池的建立尚未得到解决。目前缺乏适当的遗传工具来研究 HSC 体内克隆水平一直是一个主要障碍。我们的新型遗传工具将允许在克隆水平上追踪 HSC 体内。如果成功，研究结果将直接解决成人骨骼如何生长的问题，从而显着推进该领域的发展。 骨髓HSC池建立。此外，我们提出的新技术将广泛适用于各种 造血以外的研究领域来解决基本问题。