This invention relates to methods and compositions for designing novel
fluorescent proteins, preferably to a green fluorescent proteins (GFP).
The engineered GFPs are modified by substituting negatively charged amino
acids with positively charged amino acids on the exterior of the protein
making the protein cell permeable. The ability of the engineered
fluorescent proteins to permeate cells obviates the need for
transfections, allowing these novel proteins to be used in numerous
biological applications.