Transduction of proteins into intact neutrophils.
Neutrophils and related phagocytic leukocytes are notoriously difficult to transfect, making the introduction of proteins into these cells for biological studies problematic. We describe here two methods that have been successfully used to introduce proteins into intact primary human neutrophils while maintaining normal functional responses. The first utilizes a lipid-based reagent that transports proteins into intact neutrophils. This method is quick, easy, and is capable of transducing greater than 90% of the neutrophils in the population being studied. The second method involves the addition of a sequence derived from the HIV TAT protein to the protein to be introduced into the neutrophil. This requires both molecular biology to generate the initial construct as well as special procedures for protein isolation and renaturation. However, it also results in highly effective functional protein delivery into human neutrophils.
Zhao
Tieming
T
Department of Immunology, The Scripps Research Institute, La Jolla, CA, USA.
Bokoch
Gary M
GM
D015756:Genes, tat; D006801:Humans; D009504:Neutrophils / Q000378:metabolism*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D011933:Reagent Kits, Diagnostic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection / Q000379:methods*
D011506:Proteins; D011933:Reagent Kits, Diagnostic; D011993:Recombinant Fusion Proteins