Alkylation and silylation of α-fluorobenzyl anion intermediates.

Simple α-fluorobenzyl anions reacted with electrophiles such as non-activated alkyl halides and chlorotrimethylsilane. Upon treatment with LTMP as the base, fluoromethylbenzenes took part in the formation of α-monofluorobenzyl anions without stabilizing o-substituents. Furthermore, the resulting α-silyl fluoromethylbenzenes reacted with electrophiles such as acetophenone and benzaldehyde in the presence of cesium fluoride to form α-fluorobenzylated alcohols.

Application of the RNA interference technique to Xenopus embryos: Specific reduction of the ß-catenin gene products by short double-stranded RNA produced by recombinant human Dicer.

RNA interference (RNAi) is a technique for suppressing the function of specific genes and is widely used in many organisms, including yeast, nematodes, flies, plants, mice, and cultured mammalian cells. As of date, this technique has not been successfully applied to Xenopus laevis embryos. In this study, we applied RNAi to Xenopus embryos using ß-catenin as a model gene. Injection of long double-stranded RNA (dsRNA) corresponding to the 3'-untranslated region of ß-catenin mRNA into embryos induced embryonic lethality without any specific phenotype. However, injection of short dsRNA, generated from long dsRNA by treatment with recombinant human Dicer, into embryos resulted in decreased expression of endogenous ß-catenin mRNA and protein, as well as decreased Wnt signaling activity in the embryos. The decrease in ß-catenin mRNA and protein levels was observed only after mid-blastula transition. Embryos injected with short dsRNA showed a characteristic phenotype of enlarged anterior structures and loss of posterior structures. These phenotypes, as well as the increased expression of the anterior gene and decreased expression of the posterior gene, suggest that RNAi against the ß-catenin gene suppresses the "late Wnt signaling" involved in proper anterior-posterior patterning of Xenopus embryos. The effect of RNAi on Xenopus embryos was also found to be sensitive to temperature. These results strongly suggest that the RNAi technique can be applied to Xenopus embryos using short dsRNAs, appropriate temperature control, and proper selection of target genes.