TY - JOUR TI - A novel approach to genetic engineering of T-cell subsets by hematopoietic stem cell infection with a bicistronic lentivirus AU - Bogert, N. V. AU - Furkel, J. AU - Din, S. AU - Braren, I. AU - Eckstein, V. AU - Müller, J. A. AU - Uhlmann, L. AU - Katus, H. A. AU - Konstandin, M. H. T2 - Scientific Reports AB - Lentiviral modification of hematopoietic stem cells (HSCs) paved the way for in vivo experimentation and therapeutic approaches in patients with genetic disease. A disadvantage of this method is the use of a ubiquitous promoter leads not only to genetic modification of the leukocyte subset of interest e.g. T-cells, but also all other subsequent leukocyte progeny of the parent HSCs. To overcome this limitation we tested a bicistronic lentivirus, enabling subset specific modifications. Designed novel lentiviral constructs harbor a global promoter (mPGK) regulating mCherry for HSCs selection and a T-cell specific promoter upstream of eGFP. Two T-cell specific promoters were assessed: the distal Lck—(dLck) and the CD3δ-promoter. Transduced HSCs were FACS sorted by mCherry expression and transferred into sublethally irradiated C57/BL6 mice. Successful transplantation and T-cell specific expression of eGFP was monitored by peripheral blood assessment. Furthermore, recruitment response of lentiviral engineered leukocytes to the site of inflammation was tested in a peritonitis model without functional impairment. Our constructed lentivirus enables fast generation of subset specific leukocyte transgenesis as shown in T-cells in vivo and opens new opportunities to modify other HSCs derived subsets in the future. DA - 2020/08/13/ PY - 2020 DO - 10.1038/s41598-020-70793-6 DP - www.nature.com VL - 10 IS - 1 SP - 13740 LA - en SN - 2045-2322 UR - https://www.nature.com/articles/s41598-020-70793-6 Y2 - 2020/08/13/17:54:14 ER -