Assessing the effectiveness of coding and non-coding regions in antisense ribosome inhibition of gene expression in Tetrahymena.
In Tetrahymena thermophila, an "antisense ribosome" technology has been developed for inhibiting gene expression and generating novel mutants. Short segments of genes are inserted in antisense orientation into an rDNA vector in a region corresponding to an external loop of the folded rRNA. DNA segments derived from the 5'-ends of genes have proven most effective in reducing cognate gene expression. To investigate the efficacy of other genic regions, we generated Tetrahymena cell lines with antisense ribosome constructs containing 100-bp DNA segments derived from the 5'-ends, 3'-ends, and internal coding regions of two non-essential genes, granule lattice protein 1 and macronuclear histone H1. The 5'- and 3'-end constructs inhibited gene expression, but antisense ribosomes derived exclusively from coding regions had little effect.
Jacobs
Mary Ellen
ME
Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 06030, USA.
Cortezzo
Donnamarie E
DE
Klobutcher
Lawrence A
LA
D000818:Animals; D005786:Gene Expression Regulation; D016372:RNA, Antisense / Q000494:pharmacology*; D012335:RNA, Ribosomal / Q000037:antagonists & inhibitors* / Q000235:genetics; D022661:RNA, Untranslated / Q000037:antagonists & inhibitors*; D013768:Tetrahymena / Q000187:drug effects* / Q000235:genetics
D016372:RNA, Antisense; D012335:RNA, Ribosomal; D022661:RNA, Untranslated