Cleavage of cottontail rabbit papillomavirus E7 RNA with an anti-E7 ribozyme.
We have designed and constructed a plasmid (pE7RZ-1) that contains a gene for a hammerhead ribozyme that specifically cleaves cottontail rabbit papillomavirus (CRPV) E7 RNA sequences in vitro. The in vitro produced transcripts of pE7RZ-1 cleave CRPV E7 target sequences in trans at 37 degrees C. Pretreatment of pE7RZ-1 RNA with poly-L-lysine (PLL) increases the stability of the RNA to greater than 2 hours in the presence of a crude papilloma cell extract and the pE7RZ-1 RNA/PLL mixture retains catalytic activity in this system. This is the first report of a papillomavirus specific ribozyme RNA cleavage and leads the way for the design of effective papillomavirus specific ribozyme based therapies.
Department of Pathology, Pennsylvania State University, College of Medicine, Hershey 17033.
J D
JD
Wisotzkey
A
A
Krizenoskas
S
S
DiAngelo
J W
JW
Kreider
D000818:Animals; D001483:Base Sequence; D012777:Cottontail rabbit papillomavirus / Q000378:metabolism*; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010957:Plasmids; D011107:Polylysine; D016337:RNA, Catalytic / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D011817:Rabbits
C079183:E7 protein, cottontail rabbit papillomavirus; D009856:Oncogene Proteins, Viral; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012367:RNA, Viral; D011107:Polylysine