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Genetic mapping of species differences via in vitro crosses in mouse embryonic stem cells

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Abstract
Discovering the genetic changes underlying species differences is a central goal in evolutionary genetics. However, hybrid crosses between species in mammals often suffer from hybrid sterility, greatly complicating genetic mapping of trait variation across species. Here, we describe a simple, robust, and transgene-free technique to generate "in vitro crosses" in hybrid mouse embryonic stem (ES) cells by inducing random mitotic cross-overs with the drug ML216, which inhibits the DNA helicase Bloom syndrome (BLM). Starting with an interspecific F1 hybrid ES cell line between the Mus musculus laboratory mouse and Mus spretus (similar to 1.5 million years of divergence), we mapped the genetic basis of drug resistance to the antimetabolite tioguanine to a single region containing hypoxanthine-guanine phosphoribosyltransferase (Hprt) in as few as 21 d through "flow mapping" by coupling in vitro crosses with fluorescence-activated cell sorting (FACS). We also show how our platform can enable direct study of developmental variation by rederiving embryos with contribution from the recombinant ES cell lines. We demonstrate how in vitro crosses can overcome major bottlenecks in mouse complex trait genetics and address fundamental questions in evolutionary biology that are otherwise intractable through traditional breeding due to high cost, small litter sizes, and/or hybrid sterility. In doing so, we describe an experimental platform toward studying evolutionary systems biology inmouse and potentially in human and other mammals, including cross-species hybrids.
Keywords
GENOME-WIDE ASSOCIATION, COMPLEX TRAITS, COLLABORATIVE CROSS, SYSTEMS, GENETICS, HOUSE MICE, HELICASE, MUS, STERILITY, HYBRIDS, C57BL/6, interspecific hybrids, mitotic recombination, QTL mapping, genetics, evolution

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MLA
Lazzarano, Stefano, Marek Kucka, Joao PL Castro, et al. “Genetic Mapping of Species Differences via in Vitro Crosses in Mouse Embryonic Stem Cells.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 115.14 (2018): 3680–3685. Print.
APA
Lazzarano, S., Kucka, M., Castro, J. P., Naumann, R., Medina, P., Fletcher, M. N., Wombacher, R., et al. (2018). Genetic mapping of species differences via in vitro crosses in mouse embryonic stem cells. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 115(14), 3680–3685.
Chicago author-date
Lazzarano, Stefano, Marek Kucka, Joao PL Castro, Ronald Naumann, Paloma Medina, Michael NC Fletcher, Rebecka Wombacher, et al. 2018. “Genetic Mapping of Species Differences via in Vitro Crosses in Mouse Embryonic Stem Cells.” Proceedings of the National Academy of Sciences of the United States of America 115 (14): 3680–3685.
Chicago author-date (all authors)
Lazzarano, Stefano, Marek Kucka, Joao PL Castro, Ronald Naumann, Paloma Medina, Michael NC Fletcher, Rebecka Wombacher, Joost Gribnau, Tino Hochepied, Marc Van Montagu, Claude Libert, and Yingguang Frank Chan. 2018. “Genetic Mapping of Species Differences via in Vitro Crosses in Mouse Embryonic Stem Cells.” Proceedings of the National Academy of Sciences of the United States of America 115 (14): 3680–3685.
Vancouver
1.
Lazzarano S, Kucka M, Castro JP, Naumann R, Medina P, Fletcher MN, et al. Genetic mapping of species differences via in vitro crosses in mouse embryonic stem cells. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. 2018;115(14):3680–5.
IEEE
[1]
S. Lazzarano et al., “Genetic mapping of species differences via in vitro crosses in mouse embryonic stem cells,” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 115, no. 14, pp. 3680–3685, 2018.
@article{8561159,
  abstract     = {Discovering the genetic changes underlying species differences is a central goal in evolutionary genetics. However, hybrid crosses between species in mammals often suffer from hybrid sterility, greatly complicating genetic mapping of trait variation across species. Here, we describe a simple, robust, and transgene-free technique to generate "in vitro crosses" in hybrid mouse embryonic stem (ES) cells by inducing random mitotic cross-overs with the drug ML216, which inhibits the DNA helicase Bloom syndrome (BLM). Starting with an interspecific F1 hybrid ES cell line between the Mus musculus laboratory mouse and Mus spretus (similar to 1.5 million years of divergence), we mapped the genetic basis of drug resistance to the antimetabolite tioguanine to a single region containing hypoxanthine-guanine phosphoribosyltransferase (Hprt) in as few as 21 d through "flow mapping" by coupling in vitro crosses with fluorescence-activated cell sorting (FACS). We also show how our platform can enable direct study of developmental variation by rederiving embryos with contribution from the recombinant ES cell lines. We demonstrate how in vitro crosses can overcome major bottlenecks in mouse complex trait genetics and address fundamental questions in evolutionary biology that are otherwise intractable through traditional breeding due to high cost, small litter sizes, and/or hybrid sterility. In doing so, we describe an experimental platform toward studying evolutionary systems biology inmouse and potentially in human and other mammals, including cross-species hybrids.},
  author       = {Lazzarano, Stefano and Kucka, Marek and Castro, Joao PL and Naumann, Ronald and Medina, Paloma and Fletcher, Michael NC and Wombacher, Rebecka and Gribnau, Joost and Hochepied, Tino and Van Montagu, Marc and Libert, Claude and Chan, Yingguang Frank},
  issn         = {0027-8424},
  journal      = {PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA},
  keywords     = {GENOME-WIDE ASSOCIATION,COMPLEX TRAITS,COLLABORATIVE CROSS,SYSTEMS,GENETICS,HOUSE MICE,HELICASE,MUS,STERILITY,HYBRIDS,C57BL/6,interspecific hybrids,mitotic recombination,QTL mapping,genetics,evolution},
  language     = {eng},
  number       = {14},
  pages        = {3680--3685},
  title        = {Genetic mapping of species differences via in vitro crosses in mouse embryonic stem cells},
  url          = {http://dx.doi.org/10.1073/pnas.1717474115},
  volume       = {115},
  year         = {2018},
}

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