Light-controlled gene silencing in zebrafish embryos.

Functional genomic studies in zebrafish frequently use synthetic oligonucleotides called morpholinos that block RNA splicing or translation. However, the constitutive activity of these reagents limits their experimental utility. We report here the synthesis of a photoactivatable morpholino targeting the no tail (ntl) gene. This caged reagent permits spatiotemporal gene regulation in vivo and the photochemical generation of functionally mosaic organisms.

[1]  S. Ekker,et al.  Effective targeted gene ‘knockdown’ in zebrafish , 2000, Nature Genetics.

[2]  Luke G Green,et al.  A stepwise huisgen cycloaddition process: copper(I)-catalyzed regioselective "ligation" of azides and terminal alkynes. , 2002, Angewandte Chemie.

[3]  A. Miyawaki,et al.  An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  I. Dmochowski,et al.  Controlling RNA digestion by RNase H with a light-activated DNA hairpin. , 2006, Angewandte Chemie.

[5]  R. Ho,et al.  The protein product of the zebrafish homologue of the mouse T gene is expressed in nuclei of the germ ring and the notochord of the early embryo. , 1992, Development.

[6]  JohnB . Taylor,et al.  DESIGN AND SYNTHESIS OF A VERSATILE PHOTOCLEAVABLE DNA BUILDING BLOCK. APPLICATION TO PHOTOTRIGGERED HYBRIDIZATION , 1995 .

[7]  C. Kimmel,et al.  The zebrafish T-box genes no tail and spadetail are required for development of trunk and tail mesoderm and medial floor plate. , 2002, Development.

[8]  Y. Yan,et al.  Genetic interactions in zebrafish midline development. , 1997, Developmental biology.

[9]  R. Ho,et al.  Induction of muscle pioneers and floor plate is distinguished by the zebrafish no tail mutation , 1993, Cell.

[10]  Derek L. Stemple,et al.  Structure and function of the notochord: an essential organ for chordate development , 2005, Development.

[11]  C. Kimmel,et al.  Specification of cell fates at the dorsal margin of the zebrafish gastrula. , 1996, Development.

[12]  A. Mccarthy Development , 1996, Current Opinion in Neurobiology.