Sequencing of PCR amplified HBV DNA pre-c and c regions in the 2.2.15 cells and antiviral action by targeted antisense oligonucleotide directed against sequence

AIM:To study the specific inhibition of HBV gene expression by liver-targeting antisense oligonucleotide (ASON) directed against pre-c and cregious in a sequence specific manner.METHODS:According to the result of direct sequencing of PCR amplified products, a 16 mer phosphorothioate analogue of the...

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Veröffentlicht in:World journal of gastroenterology : WJG 1998-10, Vol.4 (5), p.434-436
Hauptverfasser: Zhoug, Sen, Wen, Shou-Ming, Zhang, Ding-Feng, Wang, Quan-Li, Wang, Seng-Qi, Ren, Hong
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Sprache:eng
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Zusammenfassung:AIM:To study the specific inhibition of HBV gene expression by liver-targeting antisense oligonucleotide (ASON) directed against pre-c and cregious in a sequence specific manner.METHODS:According to the result of direct sequencing of PCR amplified products, a 16 mer phosphorothioate analogue of the antisense oligonucleotide (PS-ASOn) directed against the HBV U-5-like region was synthesized and then linked with one live-targeting ligand, the galactosylated poly-L-lysine.Their effect on the expression of HBV gene was observed using the 2.2.15 cells.RESULTS:HBV DNA in the 2.2.15 cells was from HBV with surface antigen subtype ayw 1 by sequencing so that antisense oligonucleotides could bind specifically to the target sequence through base piring. Under the same experimental conditions, the inhibitory rates of PS-ASON to HBsAg and HBeAg were 70% and 58% at a concentration of 10 mol/L, while by ligand-PS-ASON they were 96% and 82%, the amount of HBV DNA in cultured supernatant and cells was reduced significantly. An unrelated sequence oligonucleotide showed no effectiveness. All the oligonucleotides had no cytotoxicity.CONCLUSION:Antisense oligonucleotides complexed by the liver-targeting ligand can be targeted to cells via asialoglycoprotein receptors, resulting in supecific inhibition of HBV gene expression and replication.
ISSN:1007-9327
2219-2840
DOI:10.3748/wjg.v4.i5.434