title: Morpholino antisense oligo inhibits trans-splicing of pre-inositol 1,4,5-trisphosphate receptor mRNA of Trypanosoma cruzi and suppresses parasite growth and infectivity.
abstract: Morpholino antisense oligos (MAOs) are used to investigate physiological gene function by inhibiting gene translation or construction of specific alternative splicing variants by blocking cis-splicing. MAOs are attractive drug candidates for viral- and bacterial-infectious disease therapy because of properties such as in vivo stability and specificity to target genes. Recently, we showed that phosphorothioate antisense oligos against Trypanosoma cruzi inositol 1,4,5-trisphosphate receptor (TcIP(3)R) mRNA inhibit the parasite host cell infection. In the present study, we identified the spliced leader (SL) acceptor of pre-TcIP(3)R mRNA and synthesized MAO, which inhibited trans-splicing of the transcript (MAO-1). MAO-1 was found to inhibit the addition of SL-RNA to pre-TcIP(3)R mRNA by real-time RT-PCR analysis. Treatment of the parasites with MAO-1 significantly impaired the growth and infectivity into host cells. These results indicate that MAO-1 is a potential novel drug for Chagas disease and that MAOs inhibiting trans-splicing can be used to investigate the physiology of trypanosomal genes leading to the development of novel drugs.
authors.affiliation: Department of Molecular and Cellular Parasitology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan. Electronic address: muneaki@juntendo.ac.jp.
authors.forename: Muneaki
authors.initials: M
authors.lastname: Hashimoto
authors.affiliation: Department of Molecular and Cellular Parasitology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan. Electronic address: tnara@juntendo.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nara
authors.affiliation: Department of Molecular and Cellular Parasitology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan. Electronic address: tmita@juntendo.ac.jp.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Mita
authors.affiliation: Laboratory for Developmental Neurobiology, RIKEN Brain Science Institute, Saitama 351-0198, Japan; Calcium Oscillation Project, International Cooperative Research Project and Solution-Oriented Research for Science and Technology, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan. Electronic address: mikosiba@brain.riken.jp.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Mikoshiba
mesh_terms: D000818:Animals; D014355:Chagas Disease / Q000469:parasitology*; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000187:drug effects* / Q000191:economics* / Q000235:genetics*; D060172:Morpholinos / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D020040:Trans-Splicing / Q000187:drug effects*; D014349:Trypanosoma cruzi / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity
chemical_list: D053496:Inositol 1,4,5-Trisphosphate Receptors; D060172:Morpholinos; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger
keywords: Inositol 1,4,5-trisphosphate receptor; Morpholino antisense oligo; Trans-splicing; Trypanosoma cruzi