CRISPR/Cas9系统的光学调控研究进展

doi:10.6023/A24060192

化学学报 ›› 2024, Vol. 82 ›› Issue (11): 1180-1192.DOI: 10.6023/A24060192 上一篇

综述

CRISPR/Cas9系统的光学调控研究进展

薛煜雯^a, 李磊^b, 李美星^b^,^*(), 沈清明^b^,^*()

a 南京邮电大学省部共建有机电子与信息显示国家重点实验室化学与生命科学学院南京 210023
b 南京邮电大学省部共建有机电子与信息显示国家重点实验室材料科学与工程学院南京 210023

投稿日期:2024-06-12 发布日期:2024-10-08
通讯作者: 李美星, 沈清明

作者简介:

薛煜雯, 南京邮电大学化学与生命科学学院在读硕士研究生, 主要从事CRISPR/Cas13a近红外肿瘤诊疗纳米平台的构建及其应用研究.

李磊, 南京邮电大学材料与科学工程学院在读硕士研究生, 主要从事光控激活型分子荧光探针的设计及肿瘤靶向诊疗应用研究.

李美星, 博士, 副教授, 硕士生导师, 2018年博士毕业于南京大学, 同年入职南京邮电大学材料科学与工程学院, 主要从事化学生物传感及光学成像、单颗粒/单细胞分析等领域的基础和应用研究.

沈清明, 博士, 教授, 博士生导师, 江苏省“333高层次人才培养工程”中青年学术技术带头人, 江苏省青蓝工程优秀教学团队负责人. 2008年获南京大学理学博士学位, 2010年至今在南京邮电大学材料科学与工程学院工作. 主要从事功能纳米材料的设计合成、生物纳米探针的组装及医学成像、光学诊疗平台构筑及面向肿瘤诊疗的应用研究.

基金资助:
江苏省自然科学基金面上项目(BK20221326); 江苏省高等学校自然科学研究重大项目(20KJA430012); 江苏高校“青蓝工程”资助

Advances in Optical Regulation of the CRISPR/Cas9 System

Yuwen Xue^a, Lei Li^b, Meixing Li^b(), Qingming Shen^b()

a State Key Laboratory of Organic Electronics and Information Displays, School of Chemistry and Life Sciences, Nanjing University of Posts & Telecommunications, Nanjing 210023
b State Key Laboratory of Organic Electronics and Information Displays, College of Materials Science & Engineering, Nanjing University of Posts & Telecommunications, Nanjing 210023

Received:2024-06-12 Published:2024-10-08
Contact: ^*E-mail: iammxli@njupt.edu.cn; iamqmshen@njupt.edu.cn
Supported by:
Natural Science Foundation of Jiangsu Province(BK20221326); Natural Science Foundation of Jiangsu Higher Education Institutions(20KJA430012); Qing Lan Project of Jiangsu Province

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摘要/Abstract

CRISPR/Cas9基因编辑系统由规律成簇的间隔短回文重复(Clustered regularly interspaced short palindromic repeats, CRISPR)序列和CRISPR-associated protein 9 (Cas9蛋白)组成, 具有结构简单、易于改造、基因编辑能力强等特点, 在基因组编辑、转录干扰、表观遗传调控等领域具有广泛的应用潜力. 尽管CRISPR/Cas9基因编辑系统在基因编辑方面具有明显优势, 但仍无法实现对基因编辑过程的精确时空控制. 另外, 对肿瘤细胞及特定病理组织的特异性还有待提升. 其潜在的脱靶现象所带来的基因毒性也会随着Cas9活性的增强而进一步加剧, 从而极大地限制了其在复杂生物系统中的应用. 因此, 制备能够精确控制多种内源性基因表达的基因编辑系统成为当前CRISPR/Cas9研究的热点. 光, 作为一种具备高时空分辨率且非侵入性的媒介, 其持续时间、位置、波长以及强度均易于调控. 光学调控作为一种CRISPR/Cas9新型时空调控策略, 因其具有毒副作用小、高时空分辨率以及实时可控等特点而备受人们关注. 光学调控策略还可以配合荧光成像、光声成像等影像技术追踪其递送过程, 能够极大降低体内基因编辑过程的控制难度和安全风险, 从而实现CRISPR系统的可视化递送与精准时空控制. 本文旨在综述近年来CRISPR/Cas9系统中采用的各种光学调控策略, 评估这些策略的优缺点, 并对CRISPR/Cas9系统中光学调控的挑战和发展前景进行展望.

关键词: CRISPR/Cas9, 光学调控, 基因编辑, 肿瘤治疗, 特异性

CRISPR/Cas9 gene editing system consists of clustered regularly interspaced short palindromic repeats (CRISPR) sequences and CRISPR-associated protein 9 (Cas9), characterized by its simple structure, easy modification, and strong gene editing ability. It has great potential for application in genome editing, transcriptional perturbation, epigenetic regulation, and other fields. Despite its significant advantages in gene editing, the CRISPR/Cas9 system fails to achieve precise spatial and temporal control over the editing process and its cell and tissue-specific recognition capability requires improvement. The potential off-target phenomenon of genotoxicity will be further aggravated with increased Cas9 activity, greatly limiting its application in complex biological systems. Therefore, developing gene editing systems capable of precisely controlling the expression of multiple endogenous genes has become a hot topic of current CRISPR/Cas9 research. Light, as a non-invasive medium with high spatiotemporal resolution, is easy to regulate in terms of duration, location, wavelength, and intensity. Optical regulation, as a novel spatiotemporal regulation strategy of CRISPR/Cas9, has attracted much attention due to its characteristics of minimal toxic side effects, high spatiotemporal resolution, and real-time controllability. Optical regulation strategies can also be used in conjunction with imaging technologies such as fluorescence imaging and photoacoustic imaging to track the delivery process, greatly reducing the difficulty and safety risks of gene editing in vivo, thereby achieving visual delivery and precise spatiotemporal control of CRISPR systems. This review aims to summarize various optical modulation strategies employed in CRISPR/Cas9 system in recent years, evaluating the advantages and disadvantages of these strategies, and provide an outlook on the challenges and prospects of optical modulation in the CRISPR/Cas9 system.

Key words: CRISPR/Cas9, optical regulation, gene editing, cancer treatment, specificity

引用此文

薛煜雯, 李磊, 李美星, 沈清明. CRISPR/Cas9系统的光学调控研究进展[J]. 化学学报, 2024, 82(11): 1180-1192.

Yuwen Xue, Lei Li, Meixing Li, Qingming Shen. Advances in Optical Regulation of the CRISPR/Cas9 System[J]. Acta Chimica Sinica, 2024, 82(11): 1180-1192.

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图/表 20

图1. 笼状Cas9的光激活示意图[20]

Figure 1. Light activation diagram of cage Cas9[20]. Copyright ? 2015, American Chemical Society

图2. 小分子光调节蛋白的机理及其CRISPR/Cas9调节中的应用[29]

Figure 2. Methods for light-modulating protein functions with small molecules and their applications on CRISPR/Cas9 modulation[29]. Copyright ?2023 SIOC, CAS, Shanghai, & WILEY-VCH GmbH

图3. CRISPR/Cas9介导的DNA编辑的光调节的示意图[31]

Figure 3. Schematic representation of the light-regulation of CRISPR/ Cas9-mediated DNA editing[31]. Copyright ? 2020 Wiley‐VCH GmbH

图4. CRISPR-plus实现Cas9介导的DNA靶向的光激活阻断[22]

Figure 4. CRISPR-plus achieves photoactivatable blockade of Cas9-mediated DNA targeting[22]. Copyright ? 2016 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim

图5. Cas9失活机制示意图[33]

Figure 5. Schematic of Cas9 deactivation mechanism[33]

图6. 构象限制型 crRNA 原理示意图[34]

Figure 6. Schematic depiction of the conformationally restricted crRNA principle[34]

图7. 基于G-四链体的CRISPR光开关系统设计示意图[35]

Figure 7. Schematic illustration of the design of the G-quadruplex-based CRISPR photoswitch system[35]

图8. 近红外光照射下pSPN介导的光调控CRISPR/Cas9基因编辑[42]

Figure 8. Schematic of pSPN-mediated delivery and photoregulation of CRISPR/Cas9 gene editing under the NIR light irradiation[42]. Copyright ? 2019 Wiley‐VCH Verlag GmbH & Co. KGaA, Weinheim

图9. HPR@CCP纳米粒的构建及其在肿瘤综合治疗中的应用[44]

图10. 用于传递HBV基因编辑的Cas9 RNP近红外光控仿生纳米平台示意图[49]

Figure 10. Schematic illustration of the NIR light-controlled biomimetic nanoplatform for the delivery of Cas9 RNP for HBV gene editing[49]. Copyright ? 2022, The Author(s)

图11. 可光激活Cas9 (paCas9)的示意图[51]

Figure 11. Schematic of the photoactivatable Cas9 (paCas9)[51]. Copyright ? 2015, Springer Nature America, Inc.

图12. 工程化的光开关SpCas9示意图[53]

Figure 12. Schematic of engineering photoswitchable SpCas9[53]

图13. CASANOVA工作原理示意图[61]

Figure 13. Schematic of CASANOVA function[61]

图14. CRISPR/Cas9系统光遗传DNA甲基化编辑平台的设计[62]

Figure 14. Design of the optogenetic DNA methylation editing platform based on the CRISPR/Cas9 system[62]. Copyright ? 2021, American Chemical Society

图15. 光诱导转录重编程系统(CaRROT)的示意图[21]

Figure 15. Schematic of light-inducible transcriptional reprogramming device (CaRROT)[21]. Copyright ? 2018, American Chemical Society

图16. FAST系统的示意图[63]

Figure 16. Schematic of the FAST system[63]

图17. 用于CRISPR/Cas9传递的SPPF-Dex NPs的设计[23]

Figure 17. Design of SPPF-Dex NPs for CRISPR/Cas9 delivery[23]. Copyright ? 2019 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim

图18. NIR-II 激光介导的CRISPR/Cas9系统的设计示意图[69]

Figure 18. Schematic illustration on the design of NIR-II laser-mediated delivery of CRISPR/Cas9 system[69]. Copyright ? 2021 Wiley‐VCH GmbH

图19. AP-RNP-F 协同基因-光热治疗策略示意图[71]

图20. 癌症免疫治疗的光热基因组编辑策略示意图[74]

Figure 20. Schematic illustration of the photothermal genome-editing strategy for cancer immunotherapy[74]. Copyright ? 2021 Wiley‐VCH GmbH

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CRISPR/Cas9系统的光学调控研究进展

Advances in Optical Regulation of the CRISPR/Cas9 System

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