Abstract

The goal of this project is to develop a transposable element-based gene transfer system and related technologies such as gene tagging for the zebrafish. We currently have limited abilities to create transgenic zebrafish and current methods do not offer researchers in this field the opportunity to create such organisms with the ease required for the full integration of this technology into their research programs. Consequently, a vital and powerful tool remains underdeveloped and underutilized, retarding the development of zebrafish into the powerful model system for biomedical research it could become. The project outlined here takes an unprecedented approach to creating a transgenic vertebrate by employing a short inverted repeat type transposable element of the mariner/Tc1 superfamily of elements.
Four specific aims are described including determining the precision of integration of Ts1 into zebrafish genomes, determining the frequency of insertional inactivation of zebrafish genes by Ts1 vectors, determining the effects of the Ts1 vector system on endogenous zebrafish transposons and determining the mechanisms of Ts1 regulation. The project relies on a host of biochemical and genetic approaches to address these specific aims.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Research Project (R01)
Project #
5R01RR006625-07
Application #
6056703
Study Section
Biological Sciences 2 (BIOL)
Program Officer
Carrington, Jill L
Project Start
1991-08-01
Project End
2001-08-31
Budget Start
1999-09-01
Budget End
2001-08-31
Support Year
7
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
University of Minnesota Twin Cities
Department
Genetics
Type
Schools of Arts and Sciences
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455

  Publications

Clark, Karl J; Geurts, Aron M; Bell, Jason B et al. (2004) Transposon vectors for gene-trap insertional mutagenesis in vertebrates. Genesis 39:225-33
Davidson, Ann E; Balciunas, Darius; Mohn, Deanna et al. (2003) Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon. Dev Biol 263:191-202
Fahrenkrug, S C; Joshi, B; Hackett Jr, P B et al. (2000) Alternative transcriptional initiation and splicing define the translational efficiencies of zebrafish mRNAs encoding eukaryotic initiation factor 4E. Differentiation 66:15-22
Kaufman, C D; Martinez-Rodriguez, G; Hackett Jr, P B (2000) Ectopic expression of c-ski disrupts gastrulation and neural patterning in zebrafish. Mech Dev 95:147-62
Izsvak, Z; Ivics, Z; Shimoda, N et al. (1999) Short inverted-repeat transposable elements in teleost fish and implications for a mechanism of their amplification. J Mol Evol 48:13-21
Fahrenkrug, S C; Dahlquist, M O; Clark, K J et al. (1999) Dynamic and tissue-specific expression of eIF4E during zebrafish embryogenesis. Differentiation 65:191-201
Caldovic, L; Agalliu, D; Hackett, P B (1999) Position-independent expression of transgenes in zebrafish. Transgenic Res 8:321-34
Essner, J J; Johnson, R G; Hackett Jr, P B (1999) Overexpression of thyroid hormone receptor alpha 1 during zebrafish embryogenesis disrupts hindbrain patterning and implicates retinoic acid receptors in the control of hox gene expression. Differentiation 65:1-11
Ivics, Z; Hackett, P B; Plasterk, R H et al. (1997) Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Cell 91:501-10
Essner, J J; Breuer, J J; Essner, R D et al. (1997) The zebrafish thyroid hormone receptor alpha 1 is expressed during early embryogenesis and can function in transcriptional repression. Differentiation 62:107-17

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