کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10759193 | 1050417 | 2013 | 4 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Short hairpin RNA induces methylation of hepatitis B virus covalently closed circular DNA in human hepatoma cells
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کلمات کلیدی
shRNARdDMPTGScccDNAcovalently closed circular DNA - DNA متصل به جنس مخالف بسته شده استRNA interference - RNA تداخل کنندهSmall interfering RNA - RNA تداخل کوچکRNAi - RNA سرکوبگر،RNA مداخلهگر، RNA خاموش کنندهshort hairpin RNA - RNA موی سر کوتاهsiRNA - siRNATGs - TG هاLentiviral vector - بردار Lentiviralpost-transcriptional gene silencing - خاموش کردن ژن post-transcriptionaltranscriptional gene silencing - خاموش کردن ژن رونویسیRNA-directed DNA Methylation - متیلاتی DNA DNA هدایت شده توسط RNAHBV - هپاتیت بhepatitis B virus - ویروس هپاتیت بی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
Small interfering RNAs not only modulate gene expression at a post-transcriptional level, but also induce transcriptional gene silencing by RNA interference-mediated heterochromatin formation and RNA-directed DNA methylation (RdDM). However, although established in plants, there have been controversies whether RdDM operates in mammals. Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) serves as a template for viral RNA transcription, and transcriptional activity of HBV cccDNA is regulated by methylation in patients with chronic HBV infection. In this study, we stably expressed short hairpin RNA (shRNA) against HBV in human hepatoma cells to determine whether shRNA induces methylation of HBV cccDNA. HepAD38 cells which permit replication of HBV under control of tetracycline-responsive promoter were transduced with lentiviral vectors which encode sh-1580, a shRNA against the hepatitis B viral protein HBx. Bisulfite sequencing PCR analysis revealed that sh-1580 induced CpG methylations at a higher rate compared to control (31.3% vs. 12.8%, p < 0.05). The sh-1580-induced CpG methylation was localized near the target sequence of sh-1580 in more than a half of the clones. Methylation-induced transcriptional suppression was confirmed by in vitro transcription assay. These results confirm the feasibility of RdDM of HBV cccDNA in human cells. Lentiviral vector-mediated transfer of shRNA may be used as a tool for novel transcriptional modulation by epigenetic modification of HBV cccDNA.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 436, Issue 2, 28 June 2013, Pages 152-155
Journal: Biochemical and Biophysical Research Communications - Volume 436, Issue 2, 28 June 2013, Pages 152-155
نویسندگان
Hyun Kyung Park, Bo Young Min, Nam Young Kim, Eun Sun Jang, Cheol Min Shin, Young Soo Park, Jin-Hyeok Hwang, Sook-Hyang Jeong, Nayoung Kim, Dong Ho Lee, Jin-Wook Kim,