School of Life Sciences, Tianjin University, Tianjin 300072, P. R. China.
Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Tianjin Key Laboratory of Function and Application of Biological Macromolecular Structures, Tianjin 300072, P. R. China.
ACS Appl Bio Mater. 2021 Mar 15;4(3):2502-2513. doi: 10.1021/acsabm.0c01465. Epub 2021 Feb 15.
Vascular endothelial growth factor (VEGF) is the key regulator in neovascular lesions. The anti-VEGF injection is a major way to relieve retinal neovascularization and treat these diseases. However, current anti-VEGF therapeutics show significant drawbacks. The reason is the inability to effectively control its therapeutic effect. Therefore, how to controllably inhibit the VEGF target is a key point for preventing angiogenesis. Here, a CRISPR-dCas9 optogenetic nanosystem was designed for the precise regulation of pathologic neovascularization. This system is composed of a light-controlled regulatory component and transcription inhibition component. They work together to controllably and effectively inhibit the target gene's VEGF. The opto-CRISPR nanosystem achieved precise regulation according to individual differences, whereby the expression and interaction of gene was activated by light. The following representative model laser-induced choroid neovascularization and oxygen-induced retinopathy were taken as examples to verify the effect of this nanosystem. The results showed that the opto-CRISPR nanosystem was more efficacious in the light control group (NV area effectively reduced by 41.54%) than in the dark control group without light treatment. This strategy for the CRISPR-optogenetic gene nanosystem led to the development of approaches for treating severe eye diseases. Besides, any target gene of interest can be designed by merely replacing the guide RNA sequences in this system, which provided a method for light-controlled gene transcriptional repression.
血管内皮生长因子 (VEGF) 是新血管病变的关键调节因子。抗 VEGF 注射是缓解视网膜新生血管化和治疗这些疾病的主要方法。然而，目前的抗 VEGF 疗法存在明显的缺陷。原因是无法有效控制其治疗效果。因此，如何控制地抑制 VEGF 靶标是预防血管生成的关键点。在这里，设计了一种 CRISPR-dCas9 光遗传学纳米系统，用于精确调节病理性新生血管化。该系统由光控调节组件和转录抑制组件组成。它们协同作用，可有效、可控地抑制靶基因 VEGF。光控 CRISPR 纳米系统根据个体差异进行精确调节，通过光激活基因的表达和相互作用。以代表性模型激光诱导脉络膜新生血管和氧诱导视网膜病变为例，验证了该纳米系统的效果。结果表明，与无光治疗的暗控组相比，光控组的光控 CRISPR 纳米系统更有效（NV 区有效减少 41.54%）。该 CRISPR-光遗传学基因纳米系统的策略为治疗严重眼部疾病的方法提供了思路。此外，通过替换该系统中的向导 RNA 序列，可以设计任何感兴趣的靶基因，为光控基因转录抑制提供了一种方法。
