G-四链体荧光探针的应用进展

doi:10.6023/cjoc202503033

摘要/Abstract

G-四链体(G-quadruplex, G4)作为富含鸟嘌呤的非经典核酸结构, 在基因组中广泛存在, 尤其是端粒、启动子以及病毒基因组中. 在脱氧核糖核酸(DNA)复制、转录和翻译等关键生命进程中起关键调控作用. G4的结构、含量或分布状态与多种疾病的发生和发展密切相关. 因此, 实现对G4的高灵敏度精准检测, 对于疾病的早期诊断与有效治疗具有极为深远的意义. 近年来, 科研人员开发了多种与G4高特异性结合的荧光配体, 这些荧光配体不仅在构建各类分析物传感平台方面展现出广泛的应用潜力, 而且凭借其独特且优异的荧光性能, 在细胞成像技术以及疾病精准诊断等前沿研究领域中发挥着重要作用. 对近年来G4荧光探针在上述领域所取得的最新突破性研究进展进行全面且系统的梳理与总结, 并基于当前的研究现状和未来的发展趋势, 对G4荧光探针的未来研究方向提出前瞻性的展望与思考, 以期为相关领域的科研工作者提供有益的参考和启示.

关键词: 荧光, G-四链体, 生物传感, 生物成像, 癌症诊疗

G-quadruplex (G4), a non-classical nucleic acid structure rich in guanine, is widely distributed in genomes, particularly in telomeres, promoters, and viral genomes. It plays a critical regulatory role in key biological processes such as deoxyribonucleic acid (DNA) replication, transcription, and translation. Aberrations in G4 structure, abundance, or distribution are closely associated with the pathogenesis and progression of multiple diseases. Therefore, achieving highly sensitive and precise detection of G4 holds paramount significance for early disease diagnosis and effective treatment. In recent years, researchers have developed various fluorescent ligands with high specificity for G4. These ligands demonstrate broad application potential in constructing analytical sensing platforms and play pivotal roles in cutting-edge fields such as cellular imaging and precision disease diagnosis due to their unique and superior fluorescence properties. The latest breakthroughs in G4 fluorescent probes across key areas including biosensing, cellular imaging, and cancer therapy are comprehensively summarized. Based on current research status and future trends, it also provides forward-looking perspectives on the development directions of G4 fluorescent probes, aiming to offer valuable references and insights for researchers in related disciplines.

Key words: fluorescence, G-quadruplex, biosensing, bioimaging, cancer diagnosis and treatment

引用此文

吕朦朦, 贾宵雪, 王建国. G-四链体荧光探针的应用进展[J]. 有机化学, 2025, 45(11): 4070-4081.

Mengmeng Lü, Xiaoxue Jia, Jianguo Wang. Progress in the Application of G-Quadruplex Fluorescent Probes[J]. Chinese Journal of Organic Chemistry, 2025, 45(11): 4070-4081.

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图/表 10

图1. (A) G-四平面结构; (B)平行G4结构;(C)反式平行G4结构; (D)混合G4结构

Figure 1. (A) G-quadruplanar structure; (B) parallel G4 structure; (C) trans-parallel G4 structure; (D) mixed G4 structure

图2. 已报道的G4响应荧光配体分子结构

Figure 2. Reported molecular structures of G4-responsive fluorescent ligands

图3. G4与配体结合的三种模式

Figure 3. Three modes of ligand combine with G4

图4. 基于G4进行金属离子检测应用实例: (A)基于G4与Zn-DIGP的特异性结合进行钾离子(K＋)的检测[31]; (B)基于T30695和ZnPPIX的特异性结合和铅离子(Pb2＋)诱导的G4组装完成铅离子(Pb2＋)的检测[34]; (C)利用T-Hg-T与G4结构稳定性差异实现汞离子(Hg2＋)的灵敏分析[36]

Figure 4. Examples of metal ion detection applications based on G4: (A) Detection of potassium ions (K＋) based on the specific binding of G4 to Zn-DIGP;[31] (B) Detection of lead ions (Pb2＋) based on the specific binding of T30695 and ZnPPIX and lead ion-induced G4 assembly;[34] (C) Sensitive analysis of mercury ions (Hg2＋) by using the difference in structural stability between T-Hg-T and G4[36] (A) Copyright (2010) American Chemical Society; (B) Copyright (2010) American Chemical Society; (C) Copyright (2014) The Royal Society of Chemistry

图5. 基于G4与ThT结合产生的荧光信号, 以及结合核酸外切酶进行TBA的检测[54]

Figure 5. Detection of thrombin aptamer based on fluorescence signal generated by G4 binding to ThT combined with exonuclease[54] Copyright (2017) Elsevier B.V.

图6. 基于G4进行核酸分析检测平台构建实例: (A) CRISPER/Cas12a与G4结合进行多种核酸目标链检测[57]; (B)基于pH调控cDNA的构型特异性地与BTMA结合进行SARS-CoV-2的定量分析[58]

Figure 6. Examples of nucleic acid analysis and detection platform construction based on G4: (A) CRISPER/Cas12a combined with G4 for multiple nucleic acid target strand detection;[57] (B) Quantitative analysis of SARS-CoV-2 based on the configuration of pH-regulated cDNA that binds specifically to BTMA[58] (A) Copyright (2024) American Chemical Society; (B) Copyright (2022) American Chemical Society

Fluorescent ligand	G4 name	G4 topology	E_x/E_m/nm	K_d/(µmol•L^－1)	References
NIRG-2	Pu22	Parallel	850/940	1.71±1.13	[19]
5a	Pu22	Parallel	575/650	1.549±0.080	[81]
5b	Pu22	Parallel	550/630	1.783±0.210	[81]
AI7	21CTA	Hybrid	389/500	1.3	[82]
IZIN-1	mt6363	Parallel	538/660	—	[89]
ISAP	c-Myc	Parallel	640/702	1.09	[92]
SF3	mt6363	Parallel	550/650	3.5	[93]

表1. 部分G4探针的光物理性质

Table 1. Photophysical properties of some G4 probes

图7. 基于G4进行细胞成像平台构建的实例: (A) SCY-5与不同G4和核酸链结合后荧光响应效果[78]; (B) NHCouI的合成路线及其与G4结合机制[79]; (C) IZIN-1分子结构及不同G4和核酸链结合后的荧光响应效果[89]

Figure 7. Examples of cell imaging platform construction based on G4: (A) Fluorescence response effect of SCY-5 after binding to different G4 and nucleic acid chains;[78] (B) Synthesis route of NHCouI and its binding mechanism with G4;[79] (C) IZIN-1 molecular structure and fluorescence response effects of different G4 and nucleic acid chains binding[89] (A) Copyright (2024) American Chemical Society; (B) Copyright (2023) The Royal Society of Chemistry; (C) Copyright (2021) Elsevier B.V.

Name	Sequence (5'-3')
Pu27	TTATGGGGAGGGTGGGGAGGGTGGGGAAGG
Myc-2345	TGAGGGTGGGGAGGGTGGGGAA
Myc-1245	TGGGGAGGGTTTTTAGGGTGGGGA
Myc-22	TGAGGGTGGGTAGGGTGGGTAA
Pu241	TGAGGGTGGIGAGGGTGGGGAA GG
ckit1	CAGAGGGAGGGCGCTGGGAGGAGGGGCTG
ckit2	CCCCGGGCGGGCGCGAGGGGAGGGGAGGC
bcl-2	GTCGGGGCGAGGGCGGGGGAAGGAGGGCGCGGGCGGGGA
K-ras	GGGAGGGAGGGAAGGAGGGAGGGAGGGA
VEGF	CCCGGGGCGGGCCGGGGGCGGGGTCCCGGCGGGGCGGAG
HIF-1α	GCGAGGGCGGGGGAGAGGGGAGGGGCGCG

表2. 已报道的G4序列信息

Table 2. Reported G4 sequence information

图8. 基于G4进行癌症治疗方法实例. (A) TOH与G4结合进行的体内成像原理及成像图[91]. (B) NIRG-II荧光探针分子设计及用于肿瘤淋巴转移可视化监测[19]

Figure 8. Example cancer treatment methods based on G4. (A) (A) In vivo imaging principle and imaging diagram of TOH combined with G4.[91] Copyright (2024) Elsevier B.V. (B) Molecular design of NIRG-II fluorescent probe and its application in visualization of tumor lymphatic metastasis.[19] Copyright (2024) American Chemical Society

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