Inhibition of human La protein by RNA interference downregulates hepatitis B virus mRNA in 2.2.15 cells

World Journal of Gastroenterology : WJG
Qin NiLing-Ling Wu

Abstract

To investigate the role of human La protein in HBV mRNA expression. Three human La protein (hLa) specific siRNA expression cassettes (SECs) containing U6+1 promoter were prepared via one-step overlapping extension PCR. After transfection with SECs into HepG2 cells, inhibition effects on hLa expression were analyzed by semi-quantitative RT-PCR and Western blotting. Then, effective SECs were screened out and transfected into 2.2.15 cells, a stable HBV-producing cell line. HBV surface antigen (HBsAg) and e antigen (HBeAg) secretions into culture media were detected by microparticle enzyme immunoassay (MEIA) and HBs and HBe mRNA levels were analyzed by semi-quantitative RT-PCR. SEC products containing U6+1 snRNA promoter, and 3 sites of hLa mRNA specific siRNA were obtained successfully by one-step overlapping extension PCR and could be directly transfected into HepG2 cells, resulting in inhibition of La protein expression in both mRNA and protein levels, among which U6+1-hLa833 was the most efficient, which reduced 18.6-fold mRNA and 89% protein level respectively. In 2.2.15 cells, U6+1-hLa833 was also efficient on inhibition of hLa expression. Furthermore, semi-quantitative RT-PCR showed that HBs and HBe mRNA levels were signific...Continue Reading

References

Feb 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·M A SellsG Acs
Dec 19, 1995·Proceedings of the National Academy of Sciences of the United States of America·L V TsuiF V Chisari
Mar 8, 2000·Microbiology and Molecular Biology Reviews : MMBR·C Seeger, W S Mason
May 16, 2001·The Journal of Cell Biology·R J Maraia
Mar 23, 2002·Science·Thijn R BrummelkampReuven Agami
May 1, 2002·Nature Biotechnology·Cynthia P PaulDavid R Engelke
Jun 5, 2002·Annual Review of Biochemistry·Sandra L Wolin, Tommy Cedervall
Jul 12, 2002·Nature·Gregory J Hannon
Jul 18, 2002·The Journal of Biological Chemistry·Sven HorkeTilman Heise
Sep 13, 2002·Journal of Viral Hepatitis·Naazneen MoollaPatrick Arbuthnot
Dec 3, 2002·RNA·Daniela CastanottoJohn J Rossi
Feb 20, 2003·Nature Biotechnology·Jason W MyersJames E Ferrell
Jul 30, 2003·FEBS Letters·Deming GouLin Liu
Sep 19, 2003·AIDS Research and Human Retroviruses·Joseph AndersonRamesh Akkina
Dec 6, 2003·The EMBO Journal·Ghadiyaram ChakshusmathiSandra L Wolin
Dec 23, 2003·Proceedings of the National Academy of Sciences of the United States of America·Lianxing ZhengPeter G Schultz
Dec 31, 2003·Biochemical and Biophysical Research Communications·Chang-Mei LiuChih-Chuan Liang

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Citations

Feb 9, 2010·Biochimica Et Biophysica Acta·Mark A BayfieldRichard J Maraia
Nov 20, 2015·Dalton Transactions : an International Journal of Inorganic Chemistry·Martin James RossJ A Cowan
Mar 10, 2005·The Journal of Gene Medicine·Michael D MooreMyra O McClure
Oct 24, 2007·Molecular Biotechnology·Jia ZhangP Sirois
Dec 13, 2007·Pharmaceutical Research·Yong ChenRam I Mahato

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