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Efficient Transfection Strategy for the Spatiotemporal Control of Gene Expression in Zebrafish

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Abstract

Functional analyses of gene function by knockdown and expression approaches strongly enhance the genetic study of development. In vivo application of the introduction of inhibitors of gene expression, mRNA, and expression constructs in the target region make it possible to perform region- and stage-specific regulation of gene function in a simple manner. As a basic tool for the conditional regulation of gene expression in target tissue, we present methods for the efficient introduction of antisense morpholino oligonucleotide (MO), mRNA, and expression plasmid constructs into early and later stage zebrafish embryo and larva. Lipofection of a neuron-specific expression construct plasmid encoding green fluorescent protein (GFP) into optic vesicle resulted in clear GFP expression in the retinotectal pathway in hatched larva. Co-lipofection of MO and GFP mRNA to the presumptive head region resulted in brain-specific knockdown of the gene in mid-stage embryos.

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Acknowledgments

We thank Drs. M. Kobayashi and A. Thomson for critical reading of the manuscript. This research was supported by grants from RIKEN, BSI, CREST (JST) and the Special Coordination Fund to H. Okamoto and grants-in-aids to H. Okamoto from the Ministry of Education, Science, Technology, Sports and Culture of Japan.

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Authors and Affiliations

  1. Laboratory for Developmental Gene Regulation, Brain Science Institute, RIKEN (The Institute of Physical and Chemical Research), Wako, Saitama, 351-0198, Japan

    Hideki Ando & Hitoshi Okamoto

  2. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (JST), Chuo-ku, Tokyo, 103-0027, Japan

    Hideki Ando & Hitoshi Okamoto

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  1. Hideki Ando
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  2. Hitoshi Okamoto
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Correspondence to Hideki Ando.

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Ando, H., Okamoto, H. Efficient Transfection Strategy for the Spatiotemporal Control of Gene Expression in Zebrafish. Mar Biotechnol 8, 295–303 (2006). https://doi.org/10.1007/s10126-005-5138-6

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  • DOI: https://doi.org/10.1007/s10126-005-5138-6

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