title: Ruthenium-Caged Antisense Morpholinos for Regulating Gene Expression in Zebrafish Embryos.
abstract: Photochemical approaches afford high spatiotemporal control over molecular structure and function, for broad applications in materials and biological science. Here, we present the first example of a visible light responsive ruthenium-based photolinker, Ru(bipyridine)2(3-ethynylpyridine)2(RuBEP), which was reacted stoichiometrically with a 25mer DNA or morpholino (MO) oligonucleotide functionalized with 3' and 5' terminal azides, via Cu(I)-mediated [3+2] Huisgen cycloaddition reactions. RuBEP-caged circular morpholinos (Ru-MOs) targeting two early developmental zebrafish genes,chordinandnotail, were synthesized and testedin vivo. One-cell-stage zebrafish embryos microinjected with Ru-MO and incubated in the dark for 24 h developed normally, consistent with caging, whereas irradiation at 450 nm dissociated one 3-ethynylpyridine ligand (ϕ = 0.33) and uncaged the MO to achieve gene knockdown. As demonstrated, Ru photolinkers provide a versatile method for controlling structure and function of biopolymers.
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, Pennsylvania 19104, United States.
authors.forename: J C
authors.initials: JC
authors.lastname: Griepenburg
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, Pennsylvania 19104, United States.
authors.forename: T L
authors.initials: TL
authors.lastname: Rapp
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, Pennsylvania 19104, United States.
authors.forename: P J
authors.initials: PJ
authors.lastname: Carroll
authors.affiliation: Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, 37 John Morgan Building, 3620 Hamilton Walk, Philadelphia, Pennsylvania 19104, United States.
authors.forename: J
authors.initials: J
authors.lastname: Eberwine
authors.affiliation: Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, Pennsylvania 19104, United States.
authors.forename: I J
authors.initials: IJ
authors.lastname: Dmochowski