通过脂质纳米颗粒输送稳定的 CRISPR-Cas9 核糖核蛋白进行肺和肝脏编辑

IF 33.1 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Kai Chen, Hesong Han, Sheng Zhao, Bryant Xu, Boyan Yin, Atip Lawanprasert, Marena Trinidad, Benjamin W. Burgstone, Niren Murthy, Jennifer A. Doudna
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引用次数: 0

摘要

如果能够可靠地生产出高效的 RNP-LNP 复合物,那么通过脂质纳米粒子(LNP)传递簇状规则间距短回文重复(CRISPR)核糖核蛋白(RNPs)就能实现高效、低毒和可扩展的体内基因组编辑。在这里,我们从嗜热地衣芽孢杆菌(Geobacillus stearothermophilus,GeoCas9)中改造出一种恒温 Cas9,生成的 iGeoCas9 变体能够对细胞和器官进行比原生 GeoCas9 多 100 倍的基因组编辑。此外,iGeoCas9 RNP-LNP 复合物还能编辑各种类型的细胞,并在接受编码单链 DNA 模板的细胞中诱导同源定向修复。利用组织选择性 LNP 配方,我们观察到单次静脉注射 iGeoCas9 RNP-LNPs 的报告小鼠肝脏和肺部的基因组编辑水平达到 16-37%。此外,与生物可降解 LNPs 复合的 iGeoCas9 RNPs 在肺组织中编辑致病的 SFTPC 基因的平均效率为 19%,与之前使用病毒或非病毒递送策略观察到的基因组编辑水平相比有了很大提高。这些结果表明,恒温 Cas9 RNP-LNP 复合物可以扩大基因组编辑的治疗潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR–Cas9 ribonucleoprotein

Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR–Cas9 ribonucleoprotein

Lipid nanoparticle (LNP) delivery of clustered regularly interspaced short palindromic repeat (CRISPR) ribonucleoproteins (RNPs) could enable high-efficiency, low-toxicity and scalable in vivo genome editing if efficacious RNP–LNP complexes can be reliably produced. Here we engineer a thermostable Cas9 from Geobacillus stearothermophilus (GeoCas9) to generate iGeoCas9 variants capable of >100× more genome editing of cells and organs compared with the native GeoCas9 enzyme. Furthermore, iGeoCas9 RNP–LNP complexes edit a variety of cell types and induce homology-directed repair in cells receiving codelivered single-stranded DNA templates. Using tissue-selective LNP formulations, we observe genome-editing levels of 16‒37% in the liver and lungs of reporter mice that receive single intravenous injections of iGeoCas9 RNP–LNPs. In addition, iGeoCas9 RNPs complexed to biodegradable LNPs edit the disease-causing SFTPC gene in lung tissue with 19% average efficiency, representing a major improvement over genome-editing levels observed previously using viral or nonviral delivery strategies. These results show that thermostable Cas9 RNP–LNP complexes can expand the therapeutic potential of genome editing.

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来源期刊
Nature biotechnology
Nature biotechnology 工程技术-生物工程与应用微生物
CiteScore
63.00
自引率
1.70%
发文量
382
审稿时长
3 months
期刊介绍: Nature Biotechnology is a monthly journal that focuses on the science and business of biotechnology. It covers a wide range of topics including technology/methodology advancements in the biological, biomedical, agricultural, and environmental sciences. The journal also explores the commercial, political, ethical, legal, and societal aspects of this research. The journal serves researchers by providing peer-reviewed research papers in the field of biotechnology. It also serves the business community by delivering news about research developments. This approach ensures that both the scientific and business communities are well-informed and able to stay up-to-date on the latest advancements and opportunities in the field. Some key areas of interest in which the journal actively seeks research papers include molecular engineering of nucleic acids and proteins, molecular therapy, large-scale biology, computational biology, regenerative medicine, imaging technology, analytical biotechnology, applied immunology, food and agricultural biotechnology, and environmental biotechnology. In summary, Nature Biotechnology is a comprehensive journal that covers both the scientific and business aspects of biotechnology. It strives to provide researchers with valuable research papers and news while also delivering important scientific advancements to the business community.
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