Altay Halit Yusuf, Ozdemir Fatma, Afghah Ferdows, Kilinc Zeynep, Ahmadian Mehri, Tschopp Markus, Agca Cavit
Molecular Biology, Genetics and Bioengineering Program, Sabanci University, Istanbul, Turkey.
Department of Ophthalmology, Cantonal Hospital Aarau, Aarau, Switzerland.
Front Neurosci. 2022 Oct 20;16:924917. doi: 10.3389/fnins.2022.924917. eCollection 2022.
Gene editing and gene regulatory fields are continuously developing new and safer tools that move beyond the initial CRISPR/Cas9 technology. As more advanced applications are emerging, it becomes crucial to understand and establish more complex gene regulatory and editing tools for efficient gene therapy applications. Ophthalmology is one of the leading fields in gene therapy applications with more than 90 clinical trials and numerous proof-of-concept studies. The majority of clinical trials are gene replacement therapies that are ideal for monogenic diseases. Despite Luxturna's clinical success, there are still several limitations to gene replacement therapies including the size of the target gene, the choice of the promoter as well as the pathogenic alleles. Therefore, further attempts to employ novel gene regulatory and gene editing applications are crucial to targeting retinal diseases that have not been possible with the existing approaches. CRISPR-Cas9 technology opened up the door for corrective gene therapies with its gene editing properties. Advancements in CRISPR-Cas9-associated tools including base modifiers and prime editing already improved the efficiency and safety profile of base editing approaches. While base editing is a highly promising effort, gene regulatory approaches that do not interfere with genomic changes are also becoming available as safer alternatives. Antisense oligonucleotides are one of the most commonly used approaches for correcting splicing defects or eliminating mutant mRNA. More complex gene regulatory methodologies like artificial transcription factors are also another developing field that allows targeting haploinsufficiency conditions, functionally equivalent genes, and multiplex gene regulation. In this review, we summarized the novel gene editing and gene regulatory technologies and highlighted recent translational progress, potential applications, and limitations with a focus on retinal diseases.
基因编辑和基因调控领域正在不断开发超越最初的CRISPR/Cas9技术的更新、更安全的工具。随着更先进的应用不断涌现，理解并建立更复杂的基因调控和编辑工具以实现高效的基因治疗应用变得至关重要。眼科是基因治疗应用的领先领域之一，有90多项临床试验和众多概念验证研究。大多数临床试验是基因替代疗法，这对于单基因疾病来说是理想的选择。尽管Luxturna取得了临床成功，但基因替代疗法仍存在一些局限性，包括靶基因的大小、启动子的选择以及致病等位基因。因此，进一步尝试采用新型基因调控和基因编辑应用对于治疗现有方法无法治疗的视网膜疾病至关重要。CRISPR-Cas9技术凭借其基因编辑特性为矫正性基因治疗打开了大门。包括碱基编辑器和引导编辑在内的CRISPR-Cas9相关工具的进步已经提高了碱基编辑方法的效率和安全性。虽然碱基编辑是一项非常有前景的工作，但不干扰基因组变化的基因调控方法也作为更安全的替代方案出现。反义寡核苷酸是纠正剪接缺陷或消除突变mRNA最常用的方法之一。像人工转录因子这样更复杂的基因调控方法也是另一个发展中的领域，它允许针对单倍剂量不足情况、功能等效基因和多重基因调控。在这篇综述中，我们总结了新型基因编辑和基因调控技术，并重点介绍了视网膜疾病方面最近的转化进展、潜在应用和局限性。
