Controlling Circular\nRNA Encapsulation within Extracellular\nVesicles for Gene Editing and Protein Replacement

Extracellular vesicles (EVs) are a population of vesicular\nbodies\noriginating from cells, and EVs have been proven to have the potential\nto deliver different cargos, such as RNAs. However, conventional methods\nare not able to encapsulate long RNAs into EVs efficiently or may\ncompromise the integrity of EVs. In this study, we have devised a\nstrategy to encapsulate long circRNAs (>1000 nt) into EVs by harnessing\nthe sorting mechanisms of cells. This strategy utilizes the inherent\nrichness of circular RNAs in EVs and a genetic engineering method\nto increase the cytoplasmic concentration of target circRNAs, facilitating\nhighly efficient RNA back-splicing to drive the circularization of\nRNAs. This allows target circRNAs to load into EVs with high efficiency.\nFurthermore, we demonstrate the practical applications of this strategy,\nshowing that these circRNAs can be delivered by EVs to recipient cells\nfor protein expression and to mice for gene editing.