Ribozymes for specific inhibition of mRNA function in the nematode Caenorhabditis elegans.
Nine different hammerhead ribozymes were designed for three specific sites of unc-22 mRNA in C. elegans, which carry the common catalytic core and 12, 16 or 20 flanking nucleotides for base pairing with the mRNA, and tested for cleavage of short substrate RNA in vitro. All the ribozymes cleaved the substrate RNA catalytically at 37 degrees C and the activities at 37 degrees C were higher for all the ribozymes than those at 20 degrees C, the nematode growth temperature. Plasmids carrying each of a few different promoters and lacZ reporter gene were prepared and tested in the nematode as a test of vectors for the expression of ribozymes in vivo.
Kawahara
T
T
Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan.
Ohshima
Y
Y
D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology*; D029742:Caenorhabditis elegans Proteins; D002148:Calmodulin-Binding Proteins; D015801:Helminth Proteins / Q000235:genetics; D009124:Muscle Proteins / Q000235:genetics; D016337:RNA, Catalytic / Q000378:metabolism*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors*; D013379:Substrate Specificity
D029742:Caenorhabditis elegans Proteins; D002148:Calmodulin-Binding Proteins; D015801:Helminth Proteins; D009124:Muscle Proteins; D016337:RNA, Catalytic; D012333:RNA, Messenger; C489233:unc-22 protein, C elegans