Optimized nanoparticle-mediated delivery of CRISPR-Cas9 system for B cell intervention

摘要

B cells exert multiple effector functions,and dysfunctions of B cells often lead to initiation and progression of diseases,including autoimmune and inflammatory diseases.Therefore,B cell intervention may be an effective strategy to treat diseases involving B cells.The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 gene editing system has been widely used for DNA deletion,insertion,and replacement.Nanocarriers have been developed as relatively mature systems and may be applied to deliver the CRISPR-Cas9 system to B cells in vivo.In this study,we created a library of nanoparticles (NPs) with different polyethylene glycol densities and zeta potentials and screened an optimal NP for in vivo B cell targeting.The selected NP could deliver the CRISPR-Cas9 system to B cells and induce Cas9 expression inside the cell environment.Injection of the NP encapsulated with Cas9/gB220 (NPCas9/gB220) into mice could disrupt B220 expression in B cells,suggestive of its applications to intervene the expression of the target molecule in B cells.Moreover,the treatment with NPCas9/gBAFFR could decrease the number of B cells and exert therapeutic effect in rheumatoid arthritis,as B-cell activating factor receptor (BAFFR) is vital for the survival and functions of B cells.In conclusion,we developed a carrier for the delivery of the CRISPR-Cas9 gene editing system for B cell intervention that could be used for the treatment of diseases related to B cell dysfunctions.