Abstract
Adenoviral vectors primarily derived from the adenovirus serotype 5 (Ad5) are widely used for many gene transfer applications. However, they cannot efficiently infect hematopoietic cells because these cells barely express the coxsakie-adenoviral receptor (CAR). In this study, we developed a soluble fusion protein linking viral fibers and the c-Kit receptor to alter Ad5 tropism to immature hematopoietic cells. The CAR-SCF fusion protein consists of two extracellular domains of human CAR and mouse stem cell factor (SCF). CAR-SCF was added to culture of various human hematopoietic cell lines together with an Ad vector expressing the eGFP gene driven by the CMV promoter. CAR-SCF greatly enhanced Ad5-mediated gene transfer and eGFP expression in c-Kit+ cell lines. The ability of CAR-SCF to enhance Ad5 vector infectivity was dependent on cellular c-Kit expression levels. Furthermore, CAR-SCF also enhanced Ad5 vector transfection into human cord blood CD34+ cells. In conclusion, the fusion protein will allow us to efficiently retarget adenoviral vectors to c-Kit+ human immature hematopoietic cells by just adding the fusion protein to transduction culture with adenoviral vectors. This method has an advantage that all conventional Ad5 vectors can be used to infect hematopoietic cells without any reconstruction or modification of the vectors.
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