caa a22 4500 477081983 CHVBK 20180405111452.0 cr unu---uuuuu 170330e19960201xx s 000 0 eng 10.1007/BF02723038 doi (NATIONALLICENCE)springer-10.1007/BF02723038 Retroviral gene transfer in Xenopus cell lines and embryos [Elektronische Daten] [J. Burns, L. McNeill, C. Shimizu, T. Matsubara, J. Yee, T. Friedmann, B. Kurdi-Haidar, E. Maliwat, C. Holt] Summary: A new class of retroviral vector pseudotypes have an expanded host species range and can be concentrated to high titers by ultracentrifugation. These pantropic vectors contain the genome of the murine leukemia virus-based vectors and the envelope protein of vesicular stomatitis virus substituted for the amphotropic envelope protein. We tested (a) the ability of pseudotyped (pantropic) and unmodified (amphotropic) vectors to stably infect three diffeentXenopus laevis cell lines, including one derived from the embryonic retina; and (b) the ability of the concentrated pseudotyped virus to infect embryos and to mediate foreign gene expression in the embryonic CNS. Expression of the neomycin phosphotransferase gene and single copy integration of the provirus into the genome of the cell lines was demonstrated. Surprisingly, the amphotropic and pantropic vectors generated neomycin-resistant clones with similar efficiency. PCR amplification of genomic DNA from single stage 10, 20, and 25 embryos microinjected in the blastocoel or neural tube cavities with concentrated pantropic vector (108 cfu/ml) revealed proviral DNA. Microinjection of a concentrated pantropic vector containing the coding sequence for the β-galactosidase gene into the neural tube lumen of 24-h embryos yielded β-galactosidase expressing cells in the brain. Thus, retroviral vectors provide an additional approach to existing strategies for gene transfer inXenopus embryos and cell lines. Society for In Vitro Biology, 1996 CNS nationallicence β-galactosidase nationallicence neomycin phosphotransferase selection nationallicence vesicular stomatitis virus nationallicence retroviral pseudotypes nationallicence Burns J. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut McNeill L. Department of Biology, University of California San Diego, 9500 Gilman Drive, 92093-0366, La Jolla, California aut Shimizu C. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut Matsubara T. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut Yee J. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut Friedmann T. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut Kurdi-Haidar B. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut Maliwat E. Department of Biology, University of California San Diego, 9500 Gilman Drive, 92093-0366, La Jolla, California aut Holt C. Department of Biology, University of California San Diego, 9500 Gilman Drive, 92093-0366, La Jolla, California aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/2(1996-02-01), 78-84 1071-2690 32:2<78 1996 32 11626 https://doi.org/10.1007/BF02723038 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02723038 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Burns J. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut NATIONALLICENCE 700 1- McNeill L. Department of Biology, University of California San Diego, 9500 Gilman Drive, 92093-0366, La Jolla, California aut NATIONALLICENCE 700 1- Shimizu C. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut NATIONALLICENCE 700 1- Matsubara T. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut NATIONALLICENCE 700 1- Yee J. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut NATIONALLICENCE 700 1- Friedmann T. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut NATIONALLICENCE 700 1- Kurdi-Haidar B. Department of Pediatrics and Center for Molecular Genetics, UCSD, 92093, La Jolla, California aut NATIONALLICENCE 700 1- Maliwat E. Department of Biology, University of California San Diego, 9500 Gilman Drive, 92093-0366, La Jolla, California aut NATIONALLICENCE 700 1- Holt C. Department of Biology, University of California San Diego, 9500 Gilman Drive, 92093-0366, La Jolla, California aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/2(1996-02-01), 78-84 1071-2690 32:2<78 1996 32 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer