Evolution, Optimization, and Application of Genome Editing Technologies

项目总结/摘要 基因编辑技术使得能够有效地靶向引入感兴趣的所需遗传改变， 广泛的生物体，并在生物学研究和人类治疗中具有广泛的应用。首先- 第二代基因编辑平台使用位点特异性核酸内切酶（例如，RNA引导的CRISPR-Cas核酸酶） 诱导靶向DNA双链断裂，使用细胞DNA修复机制来产生突变。 最近开发的基因编辑平台包括CRISPR胞嘧啶或腺嘌呤碱基编辑器，其 使用Cas9指导的单链DNA脱氨酶来诱导靶向的C-to-T或A-to-G改变， 和CRISPR引导的prime editors（其使用依赖于逆转录酶的过程， 可以诱导任何核苷酸取代或小插入缺失突变）。虽然基因编辑技术 虽然在过去十年中，开发速度加快，但现有平台的许多重要限制仍然存在， 需要解决的问题，仍然有机会创造多种新的有影响力的技术， 预付款。对于本MIRA的下一个周期，重点将放在四个广泛重要的挑战上：1）扩大 基因编辑器脱靶检测技术的灵敏度和范围; 2）设计下一代引物 具有增强和优化架构的编辑器; 3）创建新的表观遗传编辑平台， 人类基因的可遗传激活;以及4）开发新的通用模式，用于将所有基因编辑器递送至 人体细胞

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.