Hammerhead ribozymes that selectively cleave the NPY Y1, Y4, and Y5 receptor full-length RNA.
The concept of exploiting the ribozyme catalytic center for cleaving a specific target RNA transcript was applied to the design of selective ribozymes for the rat Y1, Y4, and Y5 receptor subtypes. Ribozymes selective for the neuropeptide Y (NPY) receptor subtypes were designed and chemically modified. Recognition sites were selected both according to the extent of their sequence homology between the receptor subtypes and according to the localization within single-stranded regions accessible for hybridization. Stability of the ribozymes against nucleolytic activities was increased by introducing 2'-O-methylribonucleosides and 3'-terminal modifications, such as inverted ends or dideoxynucleosides. Ribozymes cleaving the full-length rat Y1, Y4 (1200 nt), and Y5 receptor mRNA (2200 nt) were identified. The specificity of the recognition sites and the subtype selectivity of the ribozyme-mediated cleavage was demonstrated.
Wieland
H A
HA
Department of Cardiovascular/Metabolic Research, Boehringer Ingelheim Pharma KG, Biberach, Germany.
Hamilton
B S
BS
Krist
B
B
D000818:Animals; D001483:Base Sequence; D008297:Male; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D017476:Receptors, Neuropeptide Y / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D013379:Substrate Specificity
D016337:RNA, Catalytic; D017476:Receptors, Neuropeptide Y; C097212:neuropeptide Y-Y1 receptor; C100568:neuropeptide Y5 receptor; C109491:neuropeptide Y4 receptor; D012313:RNA