Preparation of HIV TAR RNA with RNA scissors.
Two hammerhead ribozymes derived from plant pathogenic RNAs were used to cut off the HIV TAR RNA from the T7 RNA transcript through a cis cleavage reaction. Stem I of the (+)vLTSV ribozyme comprises 8 nucleotides of the 5' terminus of TAR RNA, but stem III of the (+)sTRSV ribozyme consists of 8 nucleotides of the 3' end of TAR RNA. The construct containing two GUC hammerhead ribozyme target sequences identified the cleavage sites to cut off a required RNA molecule. This method was applied for preparation of 35 nt long TAR RNA. Its activity was proved by the complex formation with the Tat protein. It seems that this approach based on RNA scissors can also be used for the generation of required RNA molecules, RNA decoys or RNA aptamers in vivo.
Wyszko
E
E
Institute of Bioorganic Chemistry of the Polish Academy of Sciences, Noskowskiego 12, 61704, Poznan, Poland.
Fuerste
J P
JP
Barciszewska
M
M
Szymanski
M
M
Adamiak
R
R
Erdmann
V A
VA
Barciszewski
J
J
D017123:Bacteriophage T7 / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D004591:Electrophoresis, Polyacrylamide Gel; D015696:Gene Products, tat / Q000235:genetics* / Q000378:metabolism*; D015497:HIV-1 / Q000737:chemistry* / Q000235:genetics*; D008957:Models, Genetic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000494:pharmacology*; D012367:RNA, Viral / Q000302:isolation & purification* / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics*; D013816:Thermodynamics; D015533:Transcriptional Activation; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D016337:RNA, Catalytic; D012367:RNA, Viral; D054322:tat Gene Products, Human Immunodeficiency Virus