Getting large macromolecules through the plasma membrane and endosomal barriers remains a major challenge. We focus on generalizable method of delivering proteins and ribonucleoproteins (RNPs) to cells in vitro and in vivo with engineered ectosomes. We have found that this method enables active cargo loading, improves the specific activity of cargo delivery, and facilitates Gectosome purification. Using the engineered ectosomes that encapsulate Cre, Ago2, and SaCas9, we have found their ability to execute designed modifications of endogenous genes in cell lines in vitro and mouse liver tissue in vivo, paving the way toward applications of this technology for the treatment of a wide range of human diseases.
keywords
developing the strategies to produce programmable gectosomes (glycoprotein-enveloped ectosomes) for the delivery of bioactive macromolecules in vitro and in vivo to implement genome modification and enable wide-ranging research and possible therapeutic applications
