CRISPR/Cas9, a full-name regular cluster-spaced short palindrome repeat sequence/CRISPR-related protein 9, is a bacterial adaptive immune system that cleaves exogenous genetic material by RNA-guided Cas9 nuclease. Represented by the most commonly used Streptococcus pyogenes-derived SPCas9 system, SpCas9 nuclease cleaves specific DNA target sites through a gRNA-recognized PAM region of 5'-NGG-3'in the genome, resulting in non-homologous end junction (NHEJ) or homologous recombination (HR) DNA repair mechanisms to achieve the editing of specific gene sites. Currently, gene therapy based on CRISPR/Cas9 has been successfully used in gene therapy of many single/polygenic genetic diseases, tumors and viral infectious diseases. More than 20 kinds of CRISPR/Cas9 related clinical treatment experiments are being carried out. However, how to efficiently deliver CRISPR/Cas9 to cells in vivo for gene editing is an urgent problem.

Professor Wang Jun's team previously developed a FDA-approved PEG-PLA or PEG-PLGA block copolymer. Cationic Lipid-Assisted Polymer Nanoparticles (CLAN) were prepared by double emulsification with cationic lipids. Granules can effectively deliver siRNA into tumor cells, T cells, macrophages and other cells to play gene silencing function. Professor Wang Jun's team has made further improvements to CRISP R/Cas9 in order to achieve efficient delivery in vivo. Firstly, the team members prepared a series of CLAN nanoparticle libraries with different characteristics and found CLAN nanoparticles which could efficiently deliver CRISPR/Cas9 into macrophages through in vivo screening. Subsequently, Professor Wang Jun's team efficiently delivered Cas9 and gRNA (mCas9/gNLRP3) targeting NLRP3 gene to macrophages of mice by screening CLAN nanoparticles. By knocking out NLRP3 gene, inhibiting the activation of NLRP3 inflammatory bodies by various inflammatory stimulants, the prevention and treatment of sepsis, peritonitis and type II diabetes in mice were successfully realized. This research has found a nano-delivery system that can efficiently deliver CRISPR/Cas9 into macrophages for gene editing. It provides a new technology for macrophage function intervention and related disease treatment. Professor Wang Jun's team is working hard to improve CRISPR/Cas9 delivery in vivo for more cells.

On October 5, 2018, the research was published online in the famous international journal Nature Communications under the title of NLRP3 inflamamsome targeting with gene editing for the differentiation of inflaminary diseases. The research was supported by the major research project of the Ministry of Science and Technology (2017YFA0250601), the project of introducing innovation and entrepreneurship team in Guangdong Province (2017ZT07S054) and the National Natural Science Foundation of China.