【交流】复旦学子报道高剂量siRNA在小鼠中可以抑制RNAi
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复旦大学生化与分子生物学实验室黄伟达所率研究小组在Biochemical Journal的九月刊上报道了如题论文。哪位大侠能下载到全文，请在DXY里共享一下，以便大家交流学习。
文摘如下：
http://www.biochemj.org/bj/390/bj3900675.htm
Biochem. J. (2005) 390 (675–679) (Printed in Great Britain)
High doses of siRNAs induce eri-1 and adar-1 gene expression and reduce the efficiency of RNA interference in the mouse
Jie HONG, Zhikang QIAN, Shuiyuan SHEN, Taishan MIN, Chang TAN, JianFeng XU, Yingchun ZHAO and Weida HUANG1
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
Key words: Escherichia-coli-expressed and enzyme-digested RNA (esiRNA), hydrodynamic injection, mouse enhanced RNA interference gene (meri-1), RNA interference (RNAi), RNA interference regulation, small interfering RNA (siRNA).
Abbreviations used: Adar, adenosine deaminase acting on RNA; CHO, Chinese-hamster ovary; dsRNA, double-stranded RNA; eri, enhanced RNA interference; esiRNA, Escherichia-coli-expressed and enzyme-digested siRNA; HBsAg, type B hepatitis virus surface antigen; HBVP, type B hepatitis virus polymerase; meri-1, mouse eri-1; NP, nucleoprotein; RDRP, RNA-dependent RNA polymerase; RISC, RNA-induced silencing complex; RNAi, RNA interference; RT, reverse transcription; siRNA, small interfering RNA.
1To whom correspondence should be addressed (email whuang@fudan.edu.cn).
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RNAi (RNA interference) is a gene-silencing mechanism that is conserved in evolution from worm to human and has been a powerful tool for gene functional research. It has been clear that the RNAi effect triggered by endogenous or exogenous siRNAs (small interfering RNAs) is transient and dose-dependent. However, there is little information on the regulation of RNAi. Recently, some proteins that regulate the RNA-silencing machinery have been identified. We have observed in previous work that the expression of target genes rebounds after being suppressed for a period of time by siRNAs. In the present study, we used secretory hepatitis B virus surface antigen gene as a reporter and compared its expression level in cell culture and mice challenged by different doses of siRNAs. A quicker and higher rebound of gene expression was observed in mice tail-vein-injected with higher doses of siRNA, and the rebound was associated with an increase in the mRNA level of meri-1 (mouse enhanced RNAi) and adar-1 (adenosine deaminase acting on RNA) genes encoding an exonuclease and RNA-specific adenosine deaminase respectively. Down-regulation of meri-1 by RNAi enhanced the sensitivity and efficiency of siRNA in inhibiting the expression of hepatitis B virus surface antigen. These results indicate that RNAi machinery may be under negative regulation, through the induction of a series of genes coding for destabilizing enzymes, by siRNAs introduced into the cell, and also suggest that a suitable amount of siRNA should be used for research or therapeutic applications.
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Received 21 April 2005/17 June 2005; accepted 11 July 2005
Published as BJ Immediate Publication 11 July 2005, doi:10.1042/BJ20050647
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The Biochemical Society, London ©2005
文摘如下：
http://www.biochemj.org/bj/390/bj3900675.htm
Biochem. J. (2005) 390 (675–679) (Printed in Great Britain)
High doses of siRNAs induce eri-1 and adar-1 gene expression and reduce the efficiency of RNA interference in the mouse
Jie HONG, Zhikang QIAN, Shuiyuan SHEN, Taishan MIN, Chang TAN, JianFeng XU, Yingchun ZHAO and Weida HUANG1
Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai 200433, China
Key words: Escherichia-coli-expressed and enzyme-digested RNA (esiRNA), hydrodynamic injection, mouse enhanced RNA interference gene (meri-1), RNA interference (RNAi), RNA interference regulation, small interfering RNA (siRNA).
Abbreviations used: Adar, adenosine deaminase acting on RNA; CHO, Chinese-hamster ovary; dsRNA, double-stranded RNA; eri, enhanced RNA interference; esiRNA, Escherichia-coli-expressed and enzyme-digested siRNA; HBsAg, type B hepatitis virus surface antigen; HBVP, type B hepatitis virus polymerase; meri-1, mouse eri-1; NP, nucleoprotein; RDRP, RNA-dependent RNA polymerase; RISC, RNA-induced silencing complex; RNAi, RNA interference; RT, reverse transcription; siRNA, small interfering RNA.
1To whom correspondence should be addressed (email whuang@fudan.edu.cn).
--------------------------------------------------------------------------------
RNAi (RNA interference) is a gene-silencing mechanism that is conserved in evolution from worm to human and has been a powerful tool for gene functional research. It has been clear that the RNAi effect triggered by endogenous or exogenous siRNAs (small interfering RNAs) is transient and dose-dependent. However, there is little information on the regulation of RNAi. Recently, some proteins that regulate the RNA-silencing machinery have been identified. We have observed in previous work that the expression of target genes rebounds after being suppressed for a period of time by siRNAs. In the present study, we used secretory hepatitis B virus surface antigen gene as a reporter and compared its expression level in cell culture and mice challenged by different doses of siRNAs. A quicker and higher rebound of gene expression was observed in mice tail-vein-injected with higher doses of siRNA, and the rebound was associated with an increase in the mRNA level of meri-1 (mouse enhanced RNAi) and adar-1 (adenosine deaminase acting on RNA) genes encoding an exonuclease and RNA-specific adenosine deaminase respectively. Down-regulation of meri-1 by RNAi enhanced the sensitivity and efficiency of siRNA in inhibiting the expression of hepatitis B virus surface antigen. These results indicate that RNAi machinery may be under negative regulation, through the induction of a series of genes coding for destabilizing enzymes, by siRNAs introduced into the cell, and also suggest that a suitable amount of siRNA should be used for research or therapeutic applications.
--------------------------------------------------------------------------------
Received 21 April 2005/17 June 2005; accepted 11 July 2005
Published as BJ Immediate Publication 11 July 2005, doi:10.1042/BJ20050647
--------------------------------------------------------------------------------
The Biochemical Society, London ©2005
