Department of PET Center, Xiangya Hospital, Central South University, Changsha, 410008, China; Laboratory of Molecular Imaging and Nanomedicine (LOMIN), National Institute of Biomedical Imaging and Bioengineering (NIBIB), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
Department of PET Center, Xiangya Hospital, Central South University, Changsha, 410008, China.
Biomaterials. 2018 Jul;171:207-218. doi: 10.1016/j.biomaterials.2018.04.031. Epub 2018 Apr 18.
In recent years, CRISPR (clustered regularly interspaced short palindromic repeat)/Cas (CRISPR-associated) genome editing systems have become one of the most robust platforms in basic biomedical research and therapeutic applications. To date, efficient in vivo delivery of the CRISPR/Cas9 system to the targeted cells remains a challenge. Although viral vectors have been widely used in the delivery of the CRISPR/Cas9 system in vitro and in vivo, their fundamental shortcomings, such as the risk of carcinogenesis, limited insertion size, immune responses and difficulty in large-scale production, severely limit their further applications. Alternative non-viral delivery systems for CRISPR/Cas9 are urgently needed. With the rapid development of non-viral vectors, lipid- or polymer-based nanocarriers have shown great potential for CRISPR/Cas9 delivery. In this review, we analyze the pros and cons of delivering CRISPR/Cas9 systems in the form of plasmid, mRNA, or protein and then discuss the limitations and challenges of CRISPR/Cas9-based genome editing. Furthermore, current non-viral vectors that have been applied for CRISPR/Cas9 delivery in vitro and in vivo are outlined in details. Finally, critical obstacles for non-viral delivery of CRISPR/Cas9 system are highlighted and promising strategies to overcome these barriers are proposed.
近年来，CRISPR（成簇规律间隔短回文重复）/Cas（CRISPR 相关）基因组编辑系统已成为基础生物医学研究和治疗应用中最强大的平台之一。迄今为止，将 CRISPR/Cas9 系统有效递送到靶向细胞仍然是一个挑战。尽管病毒载体已广泛用于 CRISPR/Cas9 系统的体外和体内递送，但它们存在致癌风险、插入大小有限、免疫反应和大规模生产困难等固有缺点，严重限制了它们的进一步应用。因此，迫切需要替代的非病毒递送系统。随着非病毒载体的快速发展，基于脂质或聚合物的纳米载体在 CRISPR/Cas9 递送上显示出巨大的潜力。在本文中，我们分析了以质粒、mRNA 或蛋白质形式递送 CRISPR/Cas9 系统的优缺点，然后讨论了基于 CRISPR/Cas9 的基因组编辑的局限性和挑战。此外，详细概述了目前已应用于 CRISPR/Cas9 体外和体内递送的非病毒载体。最后，强调了非病毒递送 CRISPR/Cas9 系统的关键障碍，并提出了克服这些障碍的有前途的策略。
