A minimised hammerhead ribozyme with activity against interleukin-2 in human cells.
A "minizyme" is a smaller version of the hammerhead ribozyme, in which stem-loop II has been replaced by a short linker. Here, we have synthesised a DNA-containing minizyme and a ribozyme, which are designed to cut within a 15-nucleotide sequence in human interleukin-2 mRNA, and have tested for their activity in vitro and in cells. In vitro at 37 degrees C, a minizyme with linker of sequence d(GTTTT) cleaves a 15-ribonucleotide synthetic substrate 5-fold slower than does the full-sized ribozyme. In human cells, the minizyme inhibits the production of interleukin-2 protein to a similar extent as does the ribozyme. Also, the minizyme and the ribozyme are more effective in cells than any of three controls: an inactive minizyme, a 15-nucleotide antisense DNA, or DNA of random sequence. The positive effect observed in cells indicates that minizymes may be useful as pharmaceuticals.
Sioud
M
M
Institute of Immunology and Rheumatology, National Hospital, Oslo, Norway.
Opstad
A
A
Hendry
P
P
Lockett
T J
TJ
Jennings
P A
PA
McCall
M J
MJ
D000818:Animals; D002460:Cell Line; D016373:DNA, Antisense; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D016337:RNA, Catalytic / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D016373:DNA, Antisense; D007376:Interleukin-2; D016337:RNA, Catalytic; D012333:RNA, Messenger