Center for Genome Editing and Recording

基因组编辑和记录中心：项目摘要 理解人类基因组的正常和病理功能，并将其翻译成 有效疗法的关键取决于确定编码的遗传信息如何赋予 表型DNA测序和生物信息学的最新进展已经提供了大量的基因组信息。 这些数据原则上是预防医学和治疗干预取得进展的关键。然而，在这方面， 实现个性化医疗的承诺将需要对DNA进行精确的询问和操作 在原位序列的规模和精度水平目前还没有。基因组编辑中心和 记录（CGER）将通过创建技术来检测，改变和记录这些挑战， 单个细胞和组织中基因组的序列和输出。基于CRISPR-Cas9基因组 CGER利用细菌的工程技术，将RNA引导的DNA切割活性与 Cas9酶与使用同源定向的双- 链断裂修复。与此同时，CGER将Cas9与DNA“碱基编辑”结构域缀合，以实现精确的 引入或校正点突变而不切割双链DNA。使用基于细胞的测定， CGER研究人员将询问人类细胞中特定的疾病相关位点，以提供新的生物学特性。 发现新的治疗靶点。总之，这些方法将使创建任何 在任何基因座上以高特异性和效率进行所需的序列改变，这对两个方面都有深远的影响。 基因组科学和实际治疗干预。为了补充这套基因组操作 技术，CGER还将开发一个高通量管道，用于测试功能基因表达 序列变异对人类疾病的影响。这条管道将识别和照亮 人类基因组序列变异、靶基因表达和与其他基因的相互作用之间的关系 基因.最后，CGER将创造新的方法来永久记录DNA中细胞状态的变化， 它们可以以单细胞RNA-seq格式读出。分子细胞记录仪的开发将集中在 主要是一个不断发展的血统追踪器，通过使命运地图的生成在前所未有的 决议，有望彻底改变正常发育和疾病进展的研究。到 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，他们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。拟议的工作建立在加州大学的能力之上， 基因组计划，哈佛大学和马萨诸塞州总医院，以创造变革性的 能力并获得最先进的研究设施和培训、教育和资源 社区外展CGER的教育和推广计划强调扩大教育渠道 吸引新的研究人员和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.