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Tetraploid embryo complementation assay has shown that mouse ES cells alone are capable of supporting embryonic development and adult life of mice. Newly established F, hybrid ES cells allow the production of ES cell-derived animals at a high enough efficiency to directly make ES cell-based genetics feasible. Here we report the establishment and characterization of 12 new F, hybrid ES cell lines and the use of one of the best (G4) in a gain- and loss-of-function genetic study, where the in vivo phenotypes were assessed directly from ES cell-derived embryos. We found the generation of G4 ES cell-derived animals to be very efficient. Furthermore, even after two consecutive rounds of genetic modifications, the majority of trans-genic lines retained the original potential of the parental lines; with 10-40% of chimeras producing ES cell-derived animals/embryos. Using these genetically altered ES cells, this success rate, in most cases, permitted the derivation of a sufficient number of mutants for initial phenotypic analyses only a few weeks after the establishment of the cell lines. Although the experimental design has to take into account a moderate level of uncontrolled damage on ES cell lines, our proof-of-principle experiment provides useful data to assist future designs harnessing the power of this technology to accelerate our understanding of gene function.
Keywords
EXCHANGE, PROTEIN, MICE, GENE, VEGF, EXPRESSION, SELECTION, hybrid, tetraploid complementation assay, vasculogenesis, ES cells

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Please use this url to cite or link to this publication:

Chicago
George, Sophia HL, Marina Gertsenstein, Kristina Vintersten, Ella Korets-Smith, John Murphy, Mary E Stevens, Jody Haigh, and Andras Nagy. 2007. “Developmental and Adult Phenotyping Directly from Mutant Embryonic Stem Cells.” Proceedings of the National Academy of Sciences of the United States of America 104 (11): 4455–4460.
APA
George, S. H., Gertsenstein, M., Vintersten, K., Korets-Smith, E., Murphy, J., Stevens, M. E., Haigh, J., et al. (2007). Developmental and adult phenotyping directly from mutant embryonic stem cells. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 104(11), 4455–4460.
Vancouver
1.
George SH, Gertsenstein M, Vintersten K, Korets-Smith E, Murphy J, Stevens ME, et al. Developmental and adult phenotyping directly from mutant embryonic stem cells. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. 2007;104(11):4455–60.
MLA
George, Sophia HL, Marina Gertsenstein, Kristina Vintersten, et al. “Developmental and Adult Phenotyping Directly from Mutant Embryonic Stem Cells.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 104.11 (2007): 4455–4460. Print.
@article{1202209,
  abstract     = {Tetraploid embryo complementation assay has shown that mouse ES cells alone are capable of supporting embryonic development and adult life of mice. Newly established F, hybrid ES cells allow the production of ES cell-derived animals at a high enough efficiency to directly make ES cell-based genetics feasible. Here we report the establishment and characterization of 12 new F, hybrid ES cell lines and the use of one of the best (G4) in a gain- and loss-of-function genetic study, where the in vivo phenotypes were assessed directly from ES cell-derived embryos. We found the generation of G4 ES cell-derived animals to be very efficient. Furthermore, even after two consecutive rounds of genetic modifications, the majority of trans-genic lines retained the original potential of the parental lines; with 10-40\% of chimeras producing ES cell-derived animals/embryos. Using these genetically altered ES cells, this success rate, in most cases, permitted the derivation of a sufficient number of mutants for initial phenotypic analyses only a few weeks after the establishment of the cell lines. Although the experimental design has to take into account a moderate level of uncontrolled damage on ES cell lines, our proof-of-principle experiment provides useful data to assist future designs harnessing the power of this technology to accelerate our understanding of gene function.},
  author       = {George, Sophia HL and Gertsenstein, Marina and Vintersten, Kristina and Korets-Smith, Ella and Murphy, John and Stevens, Mary E and Haigh, Jody and Nagy, Andras},
  issn         = {0027-8424},
  journal      = {PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA},
  keyword      = {EXCHANGE,PROTEIN,MICE,GENE,VEGF,EXPRESSION,SELECTION,hybrid,tetraploid complementation assay,vasculogenesis,ES cells},
  language     = {eng},
  number       = {11},
  pages        = {4455--4460},
  title        = {Developmental and adult phenotyping directly from mutant embryonic stem cells},
  url          = {http://dx.doi.org/10.1073/pnas.0609277104},
  volume       = {104},
  year         = {2007},
}

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