Department of Biomedical Engineering, Columbia University , New York, New York 10027, United States.
Department of Bioengineering, Rice University , Houston, Texas 77005, United States.
Chem Rev. 2017 Aug 9;117(15):9874-9906. doi: 10.1021/acs.chemrev.6b00799. Epub 2017 Jun 22.
Genome editing offers promising solutions to genetic disorders by editing DNA sequences or modulating gene expression. The clustered regularly interspaced short palindromic repeats (CRISPR)/associated protein 9 (CRISPR/Cas9) technology can be used to edit single or multiple genes in a wide variety of cell types and organisms in vitro and in vivo. Herein, we review the rapidly developing CRISPR/Cas9-based technologies for disease modeling and gene correction and recent progress toward Cas9/guide RNA (gRNA) delivery based on viral and nonviral vectors. We discuss the relative merits of delivering the genome editing elements in the form of DNA, mRNA, or protein, and the opportunities of combining viral delivery of a transgene encoding Cas9 with nonviral delivery of gRNA. We highlight the lessons learned from nonviral gene delivery in the past three decades and consider their applicability for CRISPR/Cas9 delivery. We also include a discussion of bioinformatics tools for gRNA design and chemical modifications of gRNA. Finally, we consider the extracellular and intracellular barriers to nonviral CRISPR/Cas9 delivery and propose strategies that may overcome these barriers to realize the clinical potential of CRISPR/Cas9-based genome editing.
基因组编辑通过编辑 DNA 序列或调节基因表达，为遗传疾病提供了有前景的解决方案。成簇的规律间隔的短回文重复序列（CRISPR）/相关蛋白 9（CRISPR/Cas9）技术可用于在体外和体内编辑各种细胞类型和生物体中的单个或多个基因。在此，我们综述了基于 CRISPR/Cas9 的疾病建模和基因校正技术的快速发展，以及基于病毒和非病毒载体的 Cas9/向导 RNA（gRNA）递呈的最新进展。我们讨论了以 DNA、mRNA 或蛋白形式递呈基因组编辑元件的相对优势，以及将 Cas9 编码转基因的病毒递呈与 gRNA 的非病毒递呈相结合的机会。我们强调了过去三十年中非病毒基因传递中吸取的经验教训，并考虑了它们在 CRISPR/Cas9 传递中的适用性。我们还讨论了 gRNA 设计和 gRNA 化学修饰的生物信息学工具。最后，我们考虑了非病毒 CRISPR/Cas9 传递的细胞外和细胞内障碍，并提出了可能克服这些障碍的策略，以实现基于 CRISPR/Cas9 的基因组编辑的临床潜力。
