基于mRNA的CRISPR/Cas13递送系统的体内抗病毒研究

近几年，全球大量暴发由RNA病毒感染的重大传染病，如流感病毒和新冠病毒。疫情的严重性促使全球的科学家寻找抗病毒药物，以抗击这种疾病。新冠病毒和流感病毒都能感染肺上皮细胞，造成严重肺炎。靶向沉默病毒RNA，是一种新的抗病毒基因治疗手段，本研究将从如下三个方面展开研究：1）修饰Cas13mRNA和sgRNA，以期提高CRISPR/Cas13d的组装效率和编辑效率；2）合成新的脂质体材料，寻找安全高效的脂质体递送材料和脂质体配比；3）将肺组织特异表达的细胞膜穴样内陷相关蛋白PV1的抗体偶联到LNP上，优化包装条件以期特异靶向肺组织；4）最后在小鼠体内用VP1偶联的LNP包装CRISPR/Cas13d靶向治疗感染流感病毒和新冠病毒的感染。用该系统递送CRISPR/Cas13d mRNA为治疗肺脏感染性疾病和肺脏非感染性疾病提供了新的抗病毒策略。

Recently, the emerging and remerging infectious diseases caused by RNA viruses are seriously affecting the life health of human and development of economy. Especially the pandemic of SARS-CoV-2 causing COVID-19 has infected more than 100 million people and led to more than 2.4 million deaths in the worldwide. The severity of the epidemic push scientists from all over the world to identify anti-viral strategies to combat the disease. Unfortunately, there is no effective antiviral strategy. Both SARS-CoV-2 and influenza virus infect lung epithelial cells and cause severe pneumonia. Targeted silencing RNA of virus is a novel antiviral gene therapy, this study will be carried out from the following aspects: 1) To modify Cas13 mRNA and sgRNA to improve the assembly efficiency and editing efficiency of CRISPR / Cas13d; 2) To synthesize new lipid materials to identify the safest and most efficient nano-lipoparticle delivery materials and the optimal ratio of the component of LNP; 3) To engineer Caveolae-PV1 targeting LNP to deliver CRISPR/Cas13d to lung; 4) To treat the infection of influenza and SARS-CoV-2 with the αVP1 –LNP packaged CRISPR / cas13d in mice. Using this system to deliver CRISPR/ cas13d mRNA provides a new therapeutic strategy for the treatment of pulmonary infectious diseases and non-infectious diseases in lung.