美国加州大学伯克利分校Jennifer A. Doudna等研究人员，合作通过脂质纳米粒递送稳定的CRISPR–Cas9核糖核蛋白进行肺和肝脏编辑。2024年10月16日，《自然—生物技术》杂志在线发表了这项成果。

研究人员表示，通过脂质纳米粒（LNP）递送成簇规律间隔的短回文重复序列（CRISPR）核糖核蛋白（RNP），如果能够可靠地产生有效的RNP-LNP复合物，可能实现高效、低毒且可扩展的体内基因组编辑。

研究人员嗜热脂肪地芽孢杆菌（Geobacillus stearothermophilus）中工程化了一种热稳定的Cas9（GeoCas9），生成iGeoCas9变体，其基因组编辑能力比原生GeoCas9酶高出100倍以上。此外，iGeoCas9 RNP-LNP复合物能够编辑多种细胞类型，并在接收到共递送的单链DNA模板的细胞中诱导同源定向修复。

使用组织选择性LNP制剂时，研究人员观察到报告小鼠在接受单次静脉注射iGeoCas9 RNP-LNP后，肝脏和肺中的基因组编辑水平达到16-37%。此外，与可生物降解的LNP复合的iGeoCas9 RNPs以19%的平均效率编辑了肺组织中的致病SFTPC基因，较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有显著提升。

这些结果表明，热稳定的Cas9 RNP-LNP复合物可以扩展基因组编辑的治疗潜力。

附：英文原文

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

Author: Chen, Kai, Han, Hesong, Zhao, Sheng, Xu, Bryant, Yin, Boyan, Lawanprasert, Atip, Trinidad, Marena, Burgstone, Benjamin W., Murthy, Niren, Doudna, Jennifer A.

Issue&Volume: 2024-10-16

Abstract: 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 1637% 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.

DOI: 10.1038/s41587-024-02437-3

Source: https://www.nature.com/articles/s41587-024-02437-3