Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion

Nuclear protein delivery underlies an array of biotechnological and therapeutic applications. While many variations of protein delivery methods have been described, it can still be difficult or inefficient to introduce exogenous proteins into plants. A major barrier to progress is the cell wall which is primarily composed of polysaccharides and thus only permeable to small molecules. Here, we report a partial enzymatic cell wall digestion-mediated uptake method that efficiently delivers protein into the nucleus of plant cells. Such a method allowed efficient nuclear delivery of green fluorescent protein (GFP) flanked by two nuclear localization sequences (NLS) into Arabidopsis thaliana epidermal root cells without the usual need for large doses of nanoparticles or tissue cultures. We also show that switching from daylight to far-red light-grown conditions promotes effective protein penetration into deep cell layers. This study establishes that a partial enzymatic cell wall degradation could be applied to other cell organelles by changing the localization sequence, paving the way toward the rational engineering of plants.