Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development
- Sivasubbu, S., Balciunas, D., Davidson, A.E., Pickart, M.A., Hermanson, S.B., Wangensteen, K.J., Wolbrink, D.C., and Ekker, S.C.
- Mechanisms of Development 123(7): 513-529 (Journal)
- Registered Authors
- Balciunas, Darius, Ekker, Stephen C., Hermanson, Spencer, Pickart, Michael, Sivasubbu, Sridhar
- Insertional mutagenesis, Poly(A) trap, 32 gene-trap, Gene-breaking, Sleeping Beauty, Transposon, Zebrafish, Transcription, Histone H2afz
- MeSH Terms
- Animals, Genetically Modified
- DNA Transposable Elements/genetics*
- Larva/growth & development
- Mutagenesis, Insertional*
- 16859902 Full text @ Mech. Dev.
Sivasubbu, S., Balciunas, D., Davidson, A.E., Pickart, M.A., Hermanson, S.B., Wangensteen, K.J., Wolbrink, D.C., and Ekker, S.C. (2006) Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development. Mechanisms of Development. 123(7):513-529.
We report a novel gene tagging, identification and mutagenicity ('gene-breaking') method for the zebrafish, Danio rerio. This modular approach consists of two distinct and separable molecular cassettes. The first is a gene-finding cassette. In this study, we employed a 3' gene-tagging approach that selectively 'traps' transcripts regardless of expression status, and we show that this cassette identifies both known and novel endogenous transcripts in transgenic zebrafish. The second is a transcriptional termination mutagenicity cassette assembled from a combination of a splice acceptor and polyadenylation signal to disrupt tagged transcripts upon integration into intronic sequence. We identified both novel and conserved loci as linked phenotypic mutations using this gene-breaking strategy, generating molecularly null mutations in both larval lethal and adult viable loci. We show that the Histone 2a family member z (H2afza) variant is essential for larval development through the generation of a lethal locus with a truncation of conserved carboxy-terminal residues in the protein. In principle this gene-breaking strategy is scalable for functional genomics screens and can be used in Sleeping Beauty transposon and other gene delivery systems in the zebrafish.
Genes / Markers
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Engineered Foreign Genes
Errata and Notes