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Development of antithrombotic miniribozymes that target peripheral tryptophan hydroxylase

Item Type:Article
Title:Development of antithrombotic miniribozymes that target peripheral tryptophan hydroxylase
Creators Name:Peter, J.U. and Alenina, N. and Bader, M. and Walther, D.J.
Abstract:Serotonin is not only a neurotransmitter in the central nervous system, but also a ubiquitous hormone in the periphery involved in vasoconstriction and platelet function. Tryptophan hydroxylase is the rate-limiting enzyme in serotonin biosynthesis. By gene targeting, we have shown that serotonin is synthesized independently by two different tryptophan hydroxylase isoenzymes in peripheral tissues and neurons and identified a neuronal tryptophan hydroxylase isoform. Mice deficient in peripheral tryptophan hydroxylase (TPH1) and serotonin exhibit a reduced risk of thrombosis and thromboembolism. Therefore, we designed several antitph1 hammerhead miniribozymes and tested their cleavage activity against short synthetic Tph1 RNA substrates. In vitro cleavage studies demonstrated site-specific cleavage of Tph1 mRNA that was dependent on substrate/miniribozyme ratio and duration of exposure to miniribozyme. Interestingly, we detected different in vitro cleavage rates after we had cloned the miniribozymes into tRNA expression constructs, and found one with a high cleavage rate. We also demonstrated that this active tRNA-miniribozyme chimera is capable of selectively cleaving native Tph1 mRNA in vivo, with concomitant downregulation of the serotonin biosynthesis. Therefore, this Tph1-specific miniribozyme may provide a novel and effective form of gene therapy that may be applicable to a variety of thrombotic diseases.
Keywords:TPH1, Serotonin, Thrombosis, Animals, Mice
Source:Molecular and Cellular Biochemistry
Page Range:205-215
Date:January 2007
Official Publication:https://doi.org/10.1007/s11010-006-9290-8
PubMed:View item in PubMed

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