TY - JOUR
TI - Gene transfer and genetic modification of embryonic stem cells by Cre- and Cre-PR-expressing MESV-based retroviral vectors
AU - Psarras, S.
AU - Karagianni, N.
AU - Kellendonk, C.
AU - Tronche, F.
AU - Cosset, F.-L.
AU - Stocking, C.
AU - Schirrmacher, V.
AU - von Boehmer, H.
AU - Khazaie, K.
JO - The Journal of Gene Medicine
PY - 2004
VL - 6
TODO - 1
SP - 32-42
PB - 
SN - 1099-498X, 1521-2254
TODO - 10.1002/jgm.442
TODO - adenovirus vector;  beta galactosidase;  cre recombinase;  DNA;  polyclonal antibody;  progesterone receptor;  retrovirus vector;  cre recombinase;  integrase;  progesterone receptor, animal cell;  article;  cell line;  controlled study;  DNA modification;  embryo;  embryo cell;  enzyme activity;  fibroblast;  gene induction;  gene transfer;  genetic recombination;  genetic transduction;  human;  human cell;  immunohistochemistry;  mouse;  nonhuman;  priority journal;  protein expression;  protein processing;  reporter gene;  stem cell;  target cell;  transgenics;  Vesicular stomatitis virus;  viral gene delivery system;  animal;  biosynthesis;  gene therapy;  gene transfer;  gene vector;  genetics;  methodology;  Retrovirus;  totipotent stem cell;  transgene;  transgenic mouse, Murinae;  unidentified retrovirus, Animals;  Fibroblasts;  Gene Therapy;  Gene Transfer Techniques;  Genetic Vectors;  Immunohistochemistry;  Integrases;  Mice;  Mice, Transgenic;  Receptors, Progesterone;  Retroviridae;  Totipotent Stem Cells;  Transgenes
TODO - Background. Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post-translationally inducible Cre-Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells. Methods. To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β-galactosidase only upon Cre-mediated recombination. This was used together with the ROSA26-R ES cell Cre-reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti-Cre polyclonal antibody, and by monitoring the expression of β-galactosidase. Results. Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone-inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre-PR (fusion of Cre and progesterone receptor) or β-galactosidase. Cre-mediated recombination in more than 60% of ROSA26-R ES cells was achieved when infected by a VSV-G-pseudotyped MESV retrovirus at MOI of 50. Conclusions. The MESV-based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2004 John Wiley & Sons, Ltd.
ER -