Cell Processing Center/Umbilical and Placental Cord Blood Bank (CPC/BSCUP), Bone Marrow Transplant Center (CEMO), National Cancer Institute (INCA), Rio de Janeiro, Brazil.
Oswaldo Cruz Foundation (FIOCRUZ), National Institute of Women, Children and Adolescents' Health Fernandes Figueira (IFF), Rio de Janeiro, Brazil.
Hum Gene Ther. 2024 Oct;35(19-20):781-797. doi: 10.1089/hum.2024.020.
The advent of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated nuclease 9 (Cas9) technology has revolutionized the field of genetic engineering, offering unprecedented potential for the targeted manipulation of DNA sequences. Advances in the mechanism of action of the CRISPR-Cas9 system allowed potential applicability for the treatment of genetic diseases. CRISPR-Cas9's mechanism of action involves the use of an RNA guide molecule to target-specific DNA sequences and the Cas9 enzyme to induce precise DNA cleavage. In the context of the CRISPR-Cas9 system, this review covers nonviral delivery methods for gene editing based on peptide internalization. Here, we describe critical areas of discussion such as immunogenicity, emphasizing the importance of safety, efficiency, and cost-effectiveness, particularly in the context of treating single-mutation genetic diseases using advanced editing techniques genetics as prime editor and base editor. The text discusses the versatility of cell-penetrating peptides (CPPs) in forming complexes for delivering biomolecules, particularly ribonucleoprotein for genome editing with CRISPR-Cas9 in human cells. In addition, it emphasizes the promise of combining CPPs with DNA base editing and prime editing systems. These systems, known for their simplicity and precision, hold great potential for correcting point mutations in human genetic diseases. In summary, the text provides a clear overview of the advantages of using CPPs for genome editing with CRISPR-Cas9, particularly in conjunction with advanced editing systems, highlighting their potential impact on clinical applications in the treatment of single-mutation genetic diseases. [Figure: see text].
成簇规律间隔短回文重复（CRISPR）-CRISPR 相关核酸酶 9（Cas9）技术的出现彻底改变了基因工程领域，为靶向操纵 DNA 序列提供了前所未有的潜力。CRISPR-Cas9 系统作用机制的进步使得其在治疗遗传疾病方面具有潜在适用性。CRISPR-Cas9 的作用机制涉及使用 RNA 引导分子靶向特定的 DNA 序列和 Cas9 酶诱导精确的 DNA 切割。在 CRISPR-Cas9 系统的背景下，本文综述了基于肽内化的基因编辑的非病毒递送方法。在这里，我们描述了一些关键的讨论领域，如免疫原性，强调了安全性、效率和成本效益的重要性，特别是在使用先进的编辑技术（如 Prime 编辑器和碱基编辑器）治疗单突变遗传疾病的背景下。本文讨论了细胞穿透肽（CPPs）在形成复合物以递送生物分子方面的多功能性，特别是在 CRISPR-Cas9 介导的人类细胞基因组编辑中递送核糖核蛋白。此外，还强调了将 CPP 与 DNA 碱基编辑和 Prime 编辑系统相结合的前景。这些系统以其简单性和精确性而著称，在纠正人类遗传疾病中的点突变方面具有巨大潜力。总之，本文提供了使用 CPP 进行 CRISPR-Cas9 基因组编辑的优势的清晰概述，特别是与先进的编辑系统结合使用时，强调了它们在治疗单突变遗传疾病的临床应用中的潜在影响。[图：见正文]。
