Suppression of Nonsense Mutations in Cell Culture and Mice by Multimerized Suppressor TRNA Genes

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2000 Apr 11

PMID 10757796

Citations 31

Authors

M Buvoli

A Buvoli

L A Leinwand

Affiliations

Soon will be listed here.

Abstract

We demonstrate here the first experimental suppression of a premature termination codon in vivo by using an ochre suppressor tRNA acting in an intact mouse. Multicopy tRNA expression plasmids were directly injected into skeletal muscle and into the hearts of transgenic mice carrying a reporter gene with an ochre mutation. A strategy for modulation of suppressor efficiency, applicable to diverse systems and based on tandem multimerization of the tRNA gene, is developed. The product of suppression (chloramphenicol acetyltransferase) accumulates linearly with increases in suppressor tRNA concentration to the point where the ochre-suppressing tRNA(Ser) is in four- to fivefold excess over the endogenous tRNA(Ser). The subsequent suppressor activity plateau seems to be attributable to accumulation of unmodified tRNAs. These results define many salient variables for suppression in vivo, for example, for tRNA suppression employed as gene therapy for nonsense defects.

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