Center for Genome Editing and Recording

基因组编辑和记录中心：项目总结 理解人类基因组的正常和病理功能并将其翻译的能力 对有效疗法的了解关键取决于确定编码的遗传信息是如何赋予 表型。DNA测序和生物信息学的最新进展提供了大量的基因组 这些数据在原则上掌握着预防医学和治疗干预方面取得进展的关键。然而， 实现个性化医疗的前景将需要对DNA进行准确的讯问和操作 以目前无法获得的规模和精度进行原位测序。基因组编辑和研究中心 记录(CGER)将通过创造技术来检测、更改和记录 个体细胞和组织中基因组的序列和输出。建立在CRISPR-Cas9基因组的基础上 利用来自细菌的工程技术，CGER将结合RNA引导的DNA切割活性 Cas9酶利用同源定向的双酶促DNA序列替换的策略 断股修复。同时，CGER将把Cas9与DNA“碱基编辑”结构域结合起来，以实现准确的 引入或纠正无双链DNA裂解的点突变。使用基于细胞的分析， CGER研究人员将询问人类细胞中特定的疾病相关基因，以提供新的生物学 洞察和发现新的治疗靶点。这些方法结合在一起，将能够创建任何 任何基因座上所需的序列改变都具有高特异性和高效率，对两者都有深远的影响 基因组科学和实用的治疗干预。为了补充这套基因组操作 技术方面，CGER还将开发一条高通量管道，用于测试功能基因表达 导致人类疾病的序列变异的影响。这条管道将识别和照亮 人类基因组序列变异与靶基因表达及相互作用的关系 基因。最后，CGER将创造永久记录DNA中细胞状态变化的新方法，以便 它们可以以单细胞RNA-SEQ格式读出。分子细胞记录仪的这一发展将是重点 主要是基于一种不断发展的谱系追踪，通过使命运地图能够以前所未有的速度生成 决议，有希望使对正常发育和疾病进展的研究发生革命性变化。至 为了实现其目标，该中心汇集了在功能基因组学、生物化学、 化学生物学和医学，所有这些都在开发基因组工程工具方面发挥了重要作用 跨多个系统。这项拟议的工作建立在加州大学的创新能力基础上 基因组学倡议，哈佛大学和马萨诸塞州总医院创造变革性 并获得最先进的研究设施和资源，以进行培训、教育和 社区外展。CGER的教育和推广计划强调拓宽教育渠道 并吸引新的研究人员和来自邻近领域的研究人员进行基因组学研究。

CENTER FOR GENOME EDITING AND RECORDING: PROJECT SUMMARY The ability to understand normal and pathologic functions of the human genome and to translate that knowledge into effective therapies depends critically on determining how encoded genetic information confers phenotype. Recent advances in DNA sequencing and bioinformatics have provided vast quantities of genomic data that, in principle, hold the keys to advances in preventive medicine and therapeutic intervention. However, realizing the promise of personalized medicine will require accurate interrogation and manipulation of DNA sequences in situ at a scale and level of accuracy not currently available. The Center for Genome Editing and Recording (CGER) will address these challenges by creating technologies to detect, alter and record the sequence and output of the genome in individual cells and tissues. Building on the CRISPR-Cas9 genome engineering technology harnessed from bacteria, CGER will couple the RNA-guided DNA cleavage activity of the Cas9 enzyme to strategies for enhancing DNA sequence replacement using homology-directed double- strand break repair. In parallel, CGER will conjugate Cas9 to DNA “base editing” domains to enable accurate introduction or correction of point mutations without double-stranded DNA cleavage. Using cell-based assays, CGER researchers will interrogate specific disease-associated loci in human cells to provide new biological insights and uncover new therapeutic targets. Together, these approaches will enable the creation of any desired sequence alteration at any locus with high specificity and efficiency, with profound implications for both genome science and practical therapeutic intervention. To complement this suite of genome-manipulation technologies, CGER will also develop a high-throughput pipeline for testing the functional gene expression impacts of sequence variants responsible for human disease. This pipeline will identify and illuminate the relationships between human genome sequence variations, target gene expression and interactions with other genes. Finally, CGER will create new methods for permanently recording cell state changes in DNA so that they can be read out in a single-cell RNA-seq format. This development of molecular cell recorders will focus primarily on an evolving lineage tracer that, by enabling the generation of fate maps at unprecedented resolutions, holds the promise to revolutionize studies of normal development and disease progression. To achieve its goals, the Center brings together investigators with strengths in functional genomics, biochemistry, chemical biology and medicine, all of whom have been instrumental in developing genome engineering tools across multiple systems. The work proposed builds on capabilities at the University of California, the Innovative Genomics Initiative, Harvard University and Massachusetts General Hospital to create transformative capabilities and to access state-of-the-art research facilities and resources for training, education and community outreach. CGER's Education and Outreach plan emphasizes broadening the educational pipeline and attracting new investigators and those from adjacent fields to conduct genomics research.