中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (7): 1839-1849.doi: 10.12307/2026.633

• 干细胞综述 stem cell review • 上一篇    下一篇

外泌体递送CRISPR/Cas系统在靶细胞内可实现基因编辑

王白燕1，2，杨  树1，王弋鸣1，吴梦晴3，肖  瑀4，郭梓璇1，张博艺1，冯书营1，2   

  1. 河南中医药大学，1医学院，3儿科医学院，4中医学院，河南省郑州市  450046；2河南省中医药特医食品工程研究中心，河南省郑州市  450046
  • 收稿日期:2025-02-25 修回日期:2025-06-19 接受日期:2025-07-03 出版日期:2026-03-08 发布日期:2025-08-21
  • 通讯作者: 冯书营，博士，教授，博士生导师，河南中医药大学医学院，河南省郑州市 450046；河南省中医药特医食品工程研究中心，河南省郑州市 450046
  • 作者简介:王白燕，女，1976年生，山西省定襄县人，汉族，2006年郑州大学毕业，硕士，副教授，硕士研究生导师，主要从事天然药物抗肿瘤作用机制研究。
  • 基金资助:
    河南省自然科学基金项目(232300421164)，项目负责人：冯书营；河南省高等学校重点科研项目计划基础研究专项(23ZX005)，项目负责人：冯书营；河南省科技研发计划联合基金项目(232301420070)，项目负责人：冯书营

Exosome-delivered CRISPR/Cas system enables gene editing in target cells 

Wang Baiyan1, 2, Yang Shu1, Wang Yiming1, Wu Mengqing3, Xiao Yu4, Guo Zixuan1, Zhang Boyi1, Feng Shuying1, 2   

  1. 1Medical College, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China; 2Henan Engineering Research Center of Special Medical Food of Traditional Chinese Medicine, Zhengzhou 450046, Henan Province, China; 3School of Pediatric Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China; 4School of Traditional Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China
  • Received:2025-02-25 Revised:2025-06-19 Accepted:2025-07-03 Online:2026-03-08 Published:2025-08-21
  • Contact: Feng Shuying, MD, Professor, Doctoral supervisor, Medical College, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China; Henan Engineering Research Center of Special Medical Food of Traditional Chinese Medicine, Zhengzhou 450046, Henan Province, China
  • About author:Wang Baiyan, MS, Associate professor, Master’s supervisor, Medical College, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China; Henan Engineering Research Center of Special Medical Food of Traditional Chinese Medicine, Zhengzhou 450046, Henan Province, China
  • Supported by:
    Henan Natural Science Foundation, No. 232300421164 (to FSY); Basic Research Project of Henan Provincial Key Scientific Research Project, No. 23ZX005 (to FSY); Henan Provincial Science and Technology Research and Development Joint Fund, No. 232301420070 (to FSY) 

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摘要：

文题释义：

外泌体：是细胞分泌的脂质纳米囊泡，直径在30-150 nm之间，参与细胞间物质运输等多种功能。外泌体可以装载小分子或核酸等治疗药物，然后递送到特定的细胞或组织中，实现药物的靶向递送。外泌体介导的靶向递送具有低毒性、低免疫原性和高工程改造性等优点，有望为多疾病的精准医疗带来希望。
CRISPR/Cas系统：该系统能够诱导靶向敲入或敲除突变位点，甚至是在机体或各类细胞中进行精确的序列改变，是目前应用最为广泛的基因编辑系统。

摘要
背景：CRISPR/Cas基因编辑系统具有精准基因编辑、靶向设计简单、低成本以及高效率等特性，已被广泛应用于科学研究和相关临床治疗。然而，CRISPR/Cas系统如何更安全、更高效、更精准地递送到靶细胞内，仍是一个急需解决的难题。
目的：从递送CRISPR/Cas系统的外泌体来源及工程化策略、外泌体对CRISPR/Cas系统的装载方法、CRISPR/Cas系统的生物形式及其在细胞内的作用途径等方面进行全方面综述，为该领域的研究者提供更直观、更系统的视角。
方法：以“exosome，drug delivery systems，delivery，CRISPR/Cas，gene editing，engineered exosomes，targeting”为英文检索词，以“外泌体，药物递送，CRISPR/Cas，工程化”为中文检索词，分别检索PubMed数据库及中国知网，检索时限为2014-2024年。通过仔细阅读文献的标题和摘要进行初步筛选，排除研究内容相关性差及内容重复的文献，最终纳入了78篇文献进行归纳和探讨。

结果与结论：①外泌体是一种直径30-150 nm的脂质囊泡，具有循环半衰期长、靶向组织的内在能力、良好的生物相容性以及较小的固有毒性等优势，显现出强大的靶向递送能力；②CRISPR/Cas系统虽是一种强大的基因编辑工具，然而现有的CRISPR/Cas系统递送载体各有利弊，无法完全满足需求；③递送CRISPR/Cas系统的外泌体主要来源于高产外泌体的细胞或组织，但研究者们依然需要根据研究所需进行选择或进一步工程化；④递送CRISPR/Cas系统的外泌体主要通过基因工程修饰、化学修饰、外泌体-脂质体杂交等策略实现工程化；⑤外泌体装载CRISPR/Cas系统的方法包括电穿孔、孵育、转染和主动装载途径等，选择合适的装载方法取决于CRISPR/Cas系统的理化性质；⑥外泌体递送CRISPR/Cas系统的生物形式包括质粒和核糖核蛋白复合体，两种形式各具特点，成功递送的CRISPR/Cas系统在靶细胞内实现基因编辑。

https://orcid.org/0000-0002-4102-1700(王白燕)；https://orcid.org/0000-0003-2466-4912(冯书营)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 外泌体, CRISPR/Cas系统, 基因编辑, 药物负载, 靶向递送, 工程化

Abstract: BACKGROUND: CRISPR/Cas gene editing system has been widely used in scientific research and related clinical therapy due to its characteristics of precise gene editing, simple targeted design, low cost and high efficiency. However, how to deliver CRISPR/Cas system to target cells more safely, efficiently and accurately is still an urgent problem.
OBJECTIVE: To review all aspects from the sources and engineering strategies of exosomes for delivering CRISPR/Cas systems, the loading methods of exosomes for CRISPR/Cas systems, the biological forms of CRISPR/Cas systems and their action pathways in cells, providing a more intuitive and systematic perspective for researchers in this field.
METHODS: “Exosome, drug delivery systems, delivery, CRISPR/Cas, gene editing, engineered exosomes, targeting” were used as English search terms and “exosomes, drug delivery, CRISPR/Cas, engineering” were used as Chinese search terms to search PubMed and CNKI databases, respectively. The search time range was from 2014 to 2024. Through careful reading of the title and abstract of the literature, the preliminary screening was carried out, and the literature with poor correlation and repeated content was excluded. Finally, 78 articles were included for in-depth analysis and discussion.
RESULTS AND CONCLUSION: (1) Exosomes are lipid vesicles with a diameter of 30-150 nm. They have the advantages of long circulation half-life, intrinsic ability to target tissues, good biocompatibility, and low inherent toxicity, showing strong targeted delivery ability. (2) Although CRISPR/Cas system is a powerful gene editing tool, the existing CRISPR/Cas system delivery vectors have their own advantages and disadvantages and cannot fully meet the needs. (3) Exosomes used for delivering the CRISPR/Cas system mainly originate from cells or tissues with high exosome production. However, researchers still need to select or further engineer them according to the requirements of their studies. (4) The exosomes delivering CRISPR/Cas system are mainly engineered by genetic engineering modification, chemical modification, exosome-liposome hybridization and other strategies. (5) Methods for loading the CRISPR/Cas system into exosomes include electroporation, incubation, transfection, and active loading approaches, with the choice of method depending on the physicochemical properties of the CRISPR/Cas system. (6) The biological forms of the CRISPR/Cas system delivered by exosomes include plasmids and ribonucleoprotein complexes, each with its own characteristics. CRISPR/Cas system successfully delivered by exosomes enable gene editing within target cells. 

Key words:  exosomes, CRISPR/Cas system, gene editing, drug load, targeted delivery, engineering

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