Core 2: GENETIC ENGINEERING CORE

核心2：基因工程核心

基本信息

批准号：
10238048
负责人：
J. KEITH JOUNG
金额：
$ 37.91万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10238048
关键词：
Address Area Atherosclerosis Bar Codes Biological Models Biomedical Research Cardiovascular Diseases Cardiovascular system Cell Differentiation process Cell Lineage Clustered Regularly Interspaced Short Palindromic Repeats Code Consultations DNA DNA Sequence DNA Sequence Alteration Deletion Mutation Development Disease Disease Progression Engineering Enhancers Enzymes Frequencies Gene Expression Generations Genes Genetic Diseases Genetic Engineering Genetic Transcription Genome Genome engineering Goals Guide RNA Hematological Disease Hematopoiesis Hematopoietic Hematopoietic stem cells Heritability Inflammation Insertion Mutation Investigational Therapies Journals Knowledge Length Lentivirus Link Mediating Methods Mus Mutation Myocardial Infarction Names Nonhomologous DNA End Joining Nucleic Acid Regulatory Sequences Outcome Pathway interactions Patients Process Reagent Research Research Personnel Science Scientist Services Specific qualifier value Specificity Stroke Technology Testing Therapeutic Intervention Transcriptional Regulation Untranslated RNA Variant Work base cardiovascular disorder risk design epigenome editing experimental study genome editing genome-wide genomic locus hematopoietic stem cell differentiation high risk in vivo innovation insertion/deletion mutation interest knockout gene mouse model novel nuclease programs promoter recruit repaired response stem cell proliferation translational approach

项目摘要

ABSTRACT The core objective of this Program Project is to define the relationships and interconnections between hematopoiesis (the differentiation of hematopoietic stem cells, or HSCs) and the progression of cardiovascular diseases (CVDs). To close this gap in knowledge, the Program Project will bring together field-leading researchers to investigate currently unknown factors that mediate hematopoiesis following CVD processes including myocardial infarction, atherosclerosis, and stroke, while also the dissecting contribution of these factors to CVD progression. Therefore, there exists a great opportunity to leverage the capabilities imparted by platform technologies like genome editing to engineer HSCs ex vivo and in mouse models of CVD to help address these outstanding questions. Advances in genome-editing technologies have enabled researchers to precisely alter DNA sequences and gene expression, allowing the rapid generation of biological models to test functional significance of sequence variants. Genome editing enables the permanent heritable alteration of genetic sequence by inducing targeted double-strand breaks (DSBs) in DNA at specified sequences. A complementary approach called epigenome editing leverages the DNA targeting capabilities of the nuclease platforms to transiently alter gene expression by recruiting heterologous effector domains capable of modulating transcriptional states to regulatory regions of the genome, such as promoters or enhancers, without the need for DSBs. Thus, the long-term goal of our Genome Engineering Core (Core 2) is to concurrently develop and optimize genome- and epigenome editing technologies that will enable the rapid, safe, and efficient editing of HSCs to elucidate the connection between hematopoiesis and CVD. Major objectives of Core 2 include supporting the Projects of this proposal by optimizing experimental approaches, and providing genome and epigenome editing reagents and expertise for key experiments of Projects 1 through 4. Core 2 will also develop novel and innovative genome editing, epigenome editing, and lineage tracing platforms to characterize genes, processes, and differentiation of HSCs. Specifically, Core 2 will support Projects 1-4 by enabling efficient and specific genome editing strategies in mouse HSCs, by developing novel epigenome editing reagents for use in HSCs, and by implementing and developing editing-based lineage tracing methods. Successful optimization and development of editing in mouse HSCs pursued by this Core will expedite the ability of Projects 1 through 4 to examine the relationships between hematopoiesis and CVD and more broadly, these advances will be widely applicable to the study and therapeutic intervention of cardiovascular and hematopoietic disorders, and the ex vivo genome and epigenome editing of HSCs.

抽象的该计划项目的核心目的是定义造血（造血干细胞的分化或HSC的分化）和心血管的进展疾病（CVD）。为了缩小知识的差距，该计划项目将汇集现场领先研究人员要研究当前未知因素，这些因素介导CVD过程后介导造血包括心肌梗死，动脉粥样硬化和中风，同时剖析了这些贡献 CVD进展的因素。因此，有一个很好的机会来利用赋予的能力平台技术，例如基因组编辑到工程师HSC的Ex Vivo和CVD的鼠标模型中解决这些出色的问题。基因组编辑技术的进步使研究人员能够精确改变DNA序列和基因表达，使生物模型的快速生成可以测试序列变体的功能意义。基因组编辑可以永久性地改变通过在指定序列中诱导靶向的双链断裂（DSB）来诱导靶向双链断裂（DSB），遗传序列。一个互补方法称为表观基因组编辑的核酸酶DNA靶向能力通过募集能够的异源效应子域来瞬时改变基因表达的平台将转录状态调节到基因组的调节区域，例如启动子或增强子，没有需要DSB。因此，我们的基因组工程核心（核心2）的长期目标是同时开发和优化基因组和表观基因组编辑技术，这些技术将使快速，安全和 HSC的有效编辑以阐明造血与CVD之间的联系。主要目标核心2包括通过优化实验方法来支持本提案的项目，并提供基因组和表观基因组编辑试剂和项目1至4的主要实验的专业知识。核心2将还开发出新颖而创新的基因组编辑，表观基因组编辑以及谱系跟踪平台表征HSC的基因，过程和分化。具体而言，核心2将支持项目1-4 通过开发新型表观基因组，在小鼠HSC中实现有效的特定基因组编辑策略编辑试剂用于HSC，并通过实施和开发基于编辑的谱系跟踪方法。该核心追求的鼠标HSC中编辑的成功优化和开发将加快项目1至4的能力检查造血与CVD之间的关系，更广泛地检查这些进步将广泛适用于心血管的研究和治疗干预措施造血性疾病，以及HSC的体内基因组和表观基因组编辑。