Evolution, Optimization, and Application of Genome Editing Technologies

基因组编辑技术的演进、优化与应用

• 批准号: 10330689
• 负责人: J. KEITH JOUNG
• 金额: $ 77.35万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-14 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330689
• 关键词: Address; Adenine; Architecture; Biomedical Research; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Cytosine; DNA Double Strand Break; DNA Repair; Deaminase; Detection; Engineering; Epigenetic Process; Evolution; Gene Mutation; Generations; Genes; Goals; Guide RNA; Heritability; Human; Modality; Mutation; Nucleotides; Organism; Process; RNA-Directed DNA Polymerase; Single-Stranded DNA; Site; Technology; Therapeutic; base editor; biological research; endonuclease; experimental study; gene product; genome editing; human disease; improved; insertion/deletion mutation; interest; next generation; nuclease; prime editor; technology development

Project Summary/Abstract Gene editing technologies enable the efficient targeted introduction of desired genetic alterations of interest in a wide range of organisms and have broad applications for biological research and human therapeutics. First- generation gene editing platforms use site-specific endonucleases (e.g., RNA-guided CRISPR-Cas nucleases) to induce targeted DNA double-stranded breaks, using cellular DNA repair mechanisms to create mutations. More recently developed gene editing platforms include the CRISPR cytosine or adenine base editors, which use Cas9-directed single-stranded DNA deaminases to induce targeted C-to-T or A-to-G alterations, respectively, and CRISPR-guided prime editors (which use a reverse transcriptase-dependent process that can induce any nucleotide substitution or small indel mutations). Although gene editing technology development has accelerated over the past decade, many important limitations of existing platforms have yet to be addressed and opportunities remain to create multiple new impactful technologies enabled by recent advances. For the next cycle of this MIRA, the focus will be on four broadly important Challenges: 1) expand the sensitivities and scope of gene editor off-target detection technologies; 2) engineer next-generation prime editors with enhanced and optimized architectures; 3) create new epigenetic editing platforms for targeted heritable activation of human genes; and 4) develop new universal modalities for delivery of all gene editors to human cells.

项目摘要/摘要 基因编辑技术使有效的有针对性地引入了所需的遗传改变感兴趣的遗传改变 广泛的生物，并在生物学研究和人类疗法中有广泛的应用。第一的- 生成基因编辑平台使用特定位点的内切酶（例如RNA引导的CRISPR-CAS核酸酶） 使用细胞DNA修复机制诱导靶向的DNA双链断裂，以创建突变。 最近开发的基因编辑平台包括CRISPR胞嘧啶或腺嘌呤基础编辑器，其中 使用CAS9定向的单链DNA脱氨酶来诱导靶向的C-T-T或A-TO-G改变， 分别和CRISPR指导的主要编辑器（使用逆转录酶的过程 可以诱导任何核苷酸取代或小型indel突变）。虽然基因编辑技术 在过去的十年中，开发加速了，现有平台的许多重要局限 要解决的问题，还有机会创建最新的多种新的有影响力的技术 进步。对于此Mira的下一个周期，重点将放在四个广泛重要的挑战上：1）扩展 基因编辑器的敏感性和范围脱靶检测技术； 2）工程师下一代Prime 具有增强和优化体系结构的编辑； 3）为目标创建新的表观遗传编辑平台 人类基因的遗传激活； 4）开发新的通用方式，用于将所有基因编辑者传递给 人类细胞。