Evolution, Optimization, and Application of Genome Editing Technologies

项目摘要/摘要 基因编辑技术能够有效地定向引入所需的感兴趣的基因改变 生物种类繁多，在生物研究和人类治疗方面有着广泛的应用。首先- 代基因编辑平台使用位点特异性内切酶(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修复机制创造突变。 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程序，它们 使用Cas9定向的单链DNA脱氨酶来诱导靶向的C-T或A-G改变， 和CRISPR指导的主要编辑(使用依赖于逆转录酶的过程， 可以引起任何核苷酸替换或小的插入突变)。虽然基因编辑技术 在过去的十年里，开发速度加快了，现有平台的许多重要限制还没有 有待解决，仍有机会创造多种新的有影响力的技术，由最近的 预付款。在本MIRA的下一个周期中，重点将放在四个广泛重要的挑战上：1)扩展 基因编辑离靶检测技术的灵敏度和适用范围；2)工程下一代总理 具有增强和优化的架构的编辑者；3)为目标用户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，将所有基因编辑交付给 人类细胞。

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.