Construction And Application Of A Near-infrared Light-controlled Gene Editing System Based On Gold Nanocages

Clustered regularly interspaced short palindromic repeats(CRISPR)was first discovered in bacteria and archaea as an adaptive immune mechanism that protects the bacteria themselves from invasion.Today,it has emerged as a powerful gene editing tool.Although CRISPR/Cas9 has shown many applications in gene mutation repair,construction of transgenic animals,high-throughput screening of genes and so on.However,we have found that its delivery efficiency is low,it is prone to missing the target,and it is difficult to regulate,which hinders the development of its therapeutic applications.Therefore,this paper proposes a combination drug strategy of gene editing therapy and chemotherapy with the help of gold nanocarriers to achieve photothermal spatiotemporal control,which provides a new idea for solving the current problems faced by gene editing therapy.In this study,we developed a novel near-infrared light-controlled gene editing bionic delivery system,which utilized gold nanocages(Au NCs)as delivery carriers,and lipoic acid derivatives containing guanidine(LA-Gu)were introduced into the Au NCs for loading Cas9 plasmid to construct a novel organo-inorganic hybrid Cas9 delivery system,termed as Au NCs-Gu.In order to achieve efficient gene therapy,we selected PLK1,a key oncogene in tumor cells,as the research target,and used Au NCs-Gu for delivering HSP70-Cas9-PLK1 plasmid,which was controlled by heat shock protein 70(HSP70).Additionally,the excellent photothermal properties of Au NCs were utilized to achieve near-infrared light regulation of Cas9 gene expression.The activation of HSP70-Cas9-PLK1 plasmid resulted in precise and efficient cutting of oncogene PLK1,thus effectively playing the role of gene therapy.Furthermore,Au NCs-Gu was capable of loading Combretastatin A4(CA4),a small molecule blood vessel blocker and which effectively played an anti-angiogenic role,thereby achieving a better therapeutic effect in collaboration with gene editing therapy.To enhance the in vivo stability and tumor targeting of the delivery system,we coated the above-mentioned near-infrared lightcontrolled gene editing system with A549 cell membrane,resulting in the formation of a more safe and effective tumor selective bionic delivery system.The system showed greater accumulation efficiency in the tumor site of mice and exhibited good inhibition of tumor growth and metastasis in mouse tumor models of non-small cell lung cancer(NSCLC).This bionic delivery system not only greatly prolonged the survival cycle of mice but also reflected high biological safety.In this paper,a multifunctional gene editing delivery system is presented,which can be controlled by near-infrared light.The system enables spatiotemporal controlled gene editing of oncogenes through near-infrared photothermal,promoting precision,safety,and efficiency of gene therapy.Additionally,the system’s characteristics enable multidrug delivery,allowing for synergistic enhancement of gene therapy through chemotherapy therapy strategies.This provides a new strategy for regulating gene editing therapy of tumors.