王钰, 黄归, 杨菡, 张学农. 非病毒载体递送CRISPR/Cas9系统的研究进展J. 药学学报, 2020,55(11): 2606-2617. doi: 10.16438/j.0513-4870.2020-0484
引用本文: 王钰, 黄归, 杨菡, 张学农. 非病毒载体递送CRISPR/Cas9系统的研究进展J. 药学学报, 2020,55(11): 2606-2617. doi: 10.16438/j.0513-4870.2020-0484
WANG Yu, HUANG Gui, YANG Han, ZHANG Xue-nong. Advances on nonviral vectors of CRISPR/Cas9 system for genome editingJ. Acta Pharmaceutica Sinica, 2020,55(11): 2606-2617. doi: 10.16438/j.0513-4870.2020-0484
Citation: WANG Yu, HUANG Gui, YANG Han, ZHANG Xue-nong. Advances on nonviral vectors of CRISPR/Cas9 system for genome editingJ. Acta Pharmaceutica Sinica, 2020,55(11): 2606-2617. doi: 10.16438/j.0513-4870.2020-0484

非病毒载体递送CRISPR/Cas9系统的研究进展

Advances on nonviral vectors of CRISPR/Cas9 system for genome editing

  • 摘要: 细菌和古细菌中发现的成簇规律间隔的短回文重复序列(CRISPR)/CRISPR相关蛋白(Cas)系统是通过CRISPR RNA引导核酸内切酶特异性断裂靶基因,借助同源重组和非同源末端连接修复基因组DNA,引入碱基序列的插入、缺失或替换,实现定向基因编辑的一个工具。它的出现为多个学科领域的基础研究带来极大的便利,也为临床治疗基因疾病提供了一种方式。两个主要功能组分只有进入细胞核才能发挥作用,由于细胞内和细胞外的多重障碍,高效递送CRISPR/Cas到靶组织或靶细胞仍是一个巨大的挑战,递送效率低限制了该技术的临床转化。多种递送方式中,非病毒载体与病毒载体、物理递送相比,具有独特的优点,许多学者投入了大量时间和精力设计性质优良的非病毒递送系统,包括阳离子脂质体、类脂纳米粒、阳离子聚合物、囊泡、金纳米粒、多肽和蛋白等。本文对CRISPR/Cas9的作用机制,基于质粒、mRNA和RNP的基因编辑实现方式、这些方式面临的困难和非病毒载体的研究进展进行了总结。

     

    Abstract: Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein system exerts genome editing effect through cleaving DNA double strands using RNA-guided endonuclease. Double-strand breaks were repaired via homology directed repair (HDR) or nonhomologous end joining (NHEJ), accompanied by insertions, deletions or replacements into the genome. As a powerful tool, CRISPR/Cas system has provided tremendous convenience for basic researches and may pave the path to treat genetic diseases and cancers. Genome editing could be achieved only when both CRISPR RNA and Cas protein are delivered into nucleus of target cell. Compared with physical and viral delivery, nonviral delivery of CRISPR/Cas system possesses unique advantages in terms of safety, loading capacity and preparation. Hence, many researchers have devoted themselves to the development of nonviral vectors with high delivery efficiency which is important for the application and translation of the promising technology. Advances on cationic liposomes, lipid like nanoparticles, cationic polymers, AuNPs, vesicles, polypeptides, proteins and so on have been made. We will give a brief introduction to the mechanism of CRISPR/Cas9, problems faced by nonviral delivery of CRISPR/Cas9 system in forms of plasmid, mRNA and protein; examples of non-viral vectors, hoping to give some hints on design of safe and efficient nonviral vectors for genome editing.

     

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