期刊
NATURE METHODS
卷 8, 期 6, 页码 506-U2265出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1606
关键词
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资金
- US National Institute on Drug Abuse [DA14546]
- US National Institute of General Medical Sciences [GM63904]
- National Institute of Diabetes and Digestive and Kidney Diseases [F30DK083219, P30DK084567]
- Mayo Foundation
- Council of Scientific and Industrial Research, India [FAC002]
We describe a conditional in vivo protein-trap mutagenesis system that reveals spatiotemporal protein expression dynamics and can be used to assess gene function in the vertebrate Danio rerio. Integration of pGBT-RP2.1 (RP2), a gene-breaking transposon containing a protein trap, efficiently disrupts gene expression with >97% knockdown of normal transcript amounts and simultaneously reports protein expression for each locus. The mutant alleles are revertible in somatic tissues via Cre recombinase or splice-site-blocking morpholinos and are thus to our knowledge the first systematic conditional mutant alleles outside the mouse model. We report a collection of 350 zebrafish lines that include diverse molecular loci. RP2 integrations reveal the complexity of genomic architecture and gene function in a living organism and can provide information on protein subcellular localization. The RP2 mutagenesis system is a step toward a unified 'codex' of protein expression and direct functional annotation of the vertebrate genome.
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