title: Selective cell targeting and lineage tracing of human induced pluripotent stem cells using recombinant avian retroviruses.
abstract: Human induced pluripotent stem cells (hiPSC) differentiate into multiple cell types. Selective cell targeting is often needed for analyzing gene function by overexpressing proteins in a distinct population of hiPSC-derived cell types and for monitoring cell fate in response to stimuli. However, to date, this has not been possible, as commonly used viruses enter the hiPSC via ubiquitously expressed receptors. Here, we report for the first time the application of a heterologous avian receptor, the tumor virus receptor A (TVA), to selectively transduce TVA(+) cells in a mixed cell population. Expression of the TVA surface receptor via genetic engineering renders cells susceptible for infection by avian leucosis virus (ALV). We generated hiPSC lines with this stably integrated, ectopic TVA receptor gene that expressed the receptor while retaining pluripotency. The undifferentiated hiPSC(TVA+) as well as their differentiating progeny could be infected by recombinant ALV (so-called RCAS virus) with high efficiency. Due to incomplete receptor blocking, even sequential infection of differentiating or undifferentiated TVA(+) cells was possible. In conclusion, the TVA/RCAS system provides an efficient and gentle gene transfer system for hiPSC and extends our possibilities for selective cell targeting and lineage tracing studies.
authors.affiliation: Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.|Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.
authors.forename: Laura
authors.initials: L
authors.lastname: Hildebrand
authors.affiliation: Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.|Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Seemann
authors.affiliation: Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.|Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.|College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, Republic of Korea.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kurtz
authors.affiliation: Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.|Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.|Max Planck Institute for Molecular Genetics, Berlin, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Hecht
authors.affiliation: Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.|Helmholtz-Zentrum Geesthacht (HZG), Institute of Biomaterial Science, Teltow, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Contzen
authors.affiliation: Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.|Helmholtz-Zentrum Geesthacht (HZG), Institute of Biomaterial Science, Teltow, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Gossen
authors.affiliation: Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany. harald.stachelscheid@charite.de.|Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany. harald.stachelscheid@charite.de.|Berlin Institute of Health-Stem Cell Core Facility, Berlin, Germany. harald.stachelscheid@charite.de.
authors.forename: Harald
authors.initials: H
authors.lastname: Stachelscheid
mesh_terms: D030161:Avian Proteins / Q000235:genetics* / Q000378:metabolism; D001358:Avian Sarcoma Viruses / Q000235:genetics* / Q000472:pathogenicity; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D005434:Flow Cytometry / Q000379:methods; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000821:virology*; D011991:Receptors, Virus / Q000235:genetics* / Q000378:metabolism
chemical_list: D030161:Avian Proteins; D004251:DNA Transposable Elements; D011991:Receptors, Virus; C088745:Tva receptor
keywords: ALV; Induced pluripotent stem cells; Lineage tracing; Selective cell targeting; TVA