Secreted Particle Information Transfer (SPIT) – A Cellular Platform forIn VivoGenetic Engineering

Abstract

ABSTRACTA multitude of tools now exist that allow us to precisely manipulate the human genome in a myriad of different ways. However, successful delivery of these tools to the cells of human patients remains a major barrier to their clinical implementation. Here we introduce a new cellular approach forin vivogenetic engineering,SecretedParticleInformationTransfer (SPIT) that utilizes human cells as delivery vectors forin vivogenetic engineering. We demonstrate the application of SPIT for cell-cell delivery of Cre recombinase and CRISPR-Cas9 enzymes, we show that genetic logic can be incorporated into SPIT and present the first demonstration of human cells as a delivery platform forin vivogenetic engineering in immunocompetent mice. We successfully applied SPIT to genetically modify multiple organs and tissue stem cellsin vivoincluding the liver, spleen, intestines, peripheral blood, and bone marrow. We anticipate that by harnessing the large packaging capacity of a human cell’s nucleus, the ability of human cells to engraft into patients’ long term and the capacity of human cells for complex genetic programming, that SPIT will become a paradigm shifting approach forin vivogenetic engineering.