Fig. 10

Schematic representation of lentiviral protein transduction. Lentiviral vectors can be manipulated to package a protein of interest (POI). This can be done either by fusing the POI to the C-terminal integrase (IN) protein of the GagPol polypeptide, or as shown in this illustration, by fusing it to the N-terminal matrix (MA) protein separated by a Pleckstrin homology domain (PH) to aid the anchorage to the plasma membrane. The integrase harbors the D64V mutation rendering the viral particles integrase-defective. By including a protease recognition site between the POI and MA protein, the POI is released in mature viral particles and delivered to the nucleus of the target cell by mechanisms currently unknown. Two possible mechanisms have been suggested: Transport of the POI within the viral core or diffusion through the cytoplasm. An optional vector RNA genome can be included in the viral particles, which in interest of CRISPR-Cas9 genome editing, could be a donor template for HDR. In this case, the lentiviral particles would carry all necessary components for CRISPR-based HDR within a single lentiviral particle