The effect of beta-tubulin-specific antisense oligonucleotide encapsulated in different cationic liposomes on the suppression [correction of supression] of intracellular L. donovani parasites in vitro.
An antisense oligonucleotide (20 mer) targeted to the parasite beta-tubulin gene and encapsulated in cationic liposomes, was used to test its antileishmanial activity in vitro. Cationic liposomes containing dioleyl trimethyl ammonium propane (DOTAP) were found to have higher antileishmanial activity (88% at 4 microM oligonucleotide) compared to two other liposomes with stearyl amine (SA) and cetyl trimethyl ammonium bromide (CTAB) as cations. Dot-blot experiments were performed to analyse the expression of beta-tubulin mRNA using beta-tubulin-specific radiolabelled DNA as a probe. When compared with their respective controls, beta-tubulin-specific gene expression was found to be diminished by treatment with a specific antisense oligonucleotide encapsulated in cationic liposomes (CTAB:DOPE) in a concentration-dependent manner. These experiments show that antisense oligonucleotides targeted to the beta-tubulin gene of Leishmania donovani inhibit beta-tubulin synthesis leading to the arrest of multiplication of intracellular parasites.
Dasgupta
Dalia
D
Divisions of Biomembrane and Genetic Engineering, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta-700032, India. biomembrane@iicb.res.in
Adhya
Samit
S
Basu
Mukul Kumar
MK
D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D001345:Autoradiography; D001483:Base Sequence; D002412:Cations; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D007893:Leishmania donovani / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D008081:Liposomes; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014343:Trypan Blue / Q000378:metabolism; D014404:Tubulin / Q000096:biosynthesis / Q000235:genetics*
D000981:Antiprotozoal Agents; D002412:Cations; D002593:Cetrimonium Compounds; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D014404:Tubulin; D014343:Trypan Blue; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; D000077286:Cetrimonium