Engineering and identifying supercharged proteins for macromolecule delivery into mammalian cells.
Supercharged proteins are a class of engineered or naturally occurring proteins with unusually high positive or negative net theoretical charge. Both supernegatively and superpositively charged proteins exhibit a remarkable ability to withstand thermally or chemically induced aggregation. Superpositively charged proteins are also able to penetrate mammalian cells. Associating cargo with these proteins, such as plasmid DNA, siRNA, or other proteins, can enable the functional delivery of these macromolecules into mammalian cells both in vitro and in vivo. The potency of functional delivery in some cases can exceed that of other current methods for macromolecule delivery, including the use of cell-penetrating peptides such as Tat and adenoviral delivery vectors. This chapter summarizes methods for engineering supercharged proteins, optimizing cell penetration, identifying naturally occurring supercharged proteins, and using these proteins for macromolecule delivery into mammalian cells.
Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
David B
DB
Thompson
James J
JJ
Cronican
David R
DR
Liu
D000818:Animals; D057846:Cell-Penetrating Peptides / Q000138:chemical synthesis / Q000302:isolation & purification / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods*; D004731:Endotoxins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances / Q000008:administration & dosage / Q000302:isolation & purification* / Q000378:metabolism; D051379:Mice; D010539:Permeability; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000138:chemical synthesis / Q000302:isolation & purification / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000302:isolation & purification / Q000378:metabolism; D055672:Static Electricity
D057846:Cell-Penetrating Peptides; D004731:Endotoxins; D046911:Macromolecular Substances; D011506:Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins