We have made important progress on the Tol2 project since the revised application and the supplemental data were submitted in 2008. The most significant advance is the demonstration of germ line transmission of remobilized Tol2 transposons following the injection of mRNA encoding Tol2 transposase in fertilized eggs insertion event (Fig. 1). We had previously shown that somatic remobilization of Tol2 transposons is very efficient;100% of injected embryos have novel reintegration events following injection of Tol2 transposase mRNA. Here, we demonstrate that the remobilization events are passed through the germ line and can produce tadpoles with novel GFP expression profiles. Figure 1 depicts the analysis of a single remobilized F2 animal (12M2r??5). Three of the four remobilization events found in 12M2r??5 have integrated into interesting gene loci (jvh, cen, fol). Outcross of other F2 remobilized frogs has indicated germ line transmission of novel re-integration events highlighted by the non-Mendelian frequencies of GFP-positive tadpoles in the F3 generation;that is, we observed a significant deviation from the expected 50% GFP-positive progeny from the outcross of remobilized frogs heterozygous for the Tol2XIG transposon. We are currently characterizing the novel integration events in the progeny from the other remobilized F2 frogs. We anticipate submitting a manuscript describing this data soon.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH079381-02
Application #
7826691
Study Section
Special Emphasis Panel (ZRG1-GGG-T (52))
Program Officer
Freund, Michelle
Project Start
2009-05-05
Project End
2012-04-30
Budget Start
2010-05-01
Budget End
2012-04-30
Support Year
2
Fiscal Year
2010
Total Cost
$437,067
Indirect Cost
Name
St. Jude Children's Research Hospital
Department
Type
DUNS #
067717892
City
Memphis
State
TN
Country
United States
Zip Code
38105
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Kelley, Clair M; Yergeau, Donald A; Zhu, Haiqing et al. (2012) Xenopus transgenics: methods using transposons. Methods Mol Biol 917:231-43
Yergeau, Donald A; Kelley, Clair M; Zhu, Haiqing et al. (2010) Transposon transgenesis in Xenopus. Methods 51:92-100
Hellsten, Uffe; Harland, Richard M; Gilchrist, Michael J et al. (2010) The genome of the Western clawed frog Xenopus tropicalis. Science 328:633-6
Yergeau, Donald A; Kelley, Clair M; Kuliyev, Emin et al. (2010) Remobilization of Tol2 transposons in Xenopus tropicalis. BMC Dev Biol 10:11
Yergeau, Donald A; Johnson Hamlet, Michelle R; Kuliyev, Emin et al. (2009) Transgenesis in Xenopus using the Sleeping Beauty transposon system. Dev Dyn 238:1727-43