Bethune International Peace Hospital

nancyling

Background & Aims
RNA interference (RNAi) may offer new treatment options for chronic hepatitis
B. Replicating via an RNA intermediate, hepatitis B virus (HBV) is known to be
principally vulnerable to RNAi. However, beyond delivery, the relevant issues
of potential off-target effects, target site conservation in circulating HBV
strains, and efficacy of RNAi itself have not systematically been addressed,
nor can the different existing data be quantitatively compared. The aim of
this study was to provide such information.
Methods
To focus on the intracellular RNAi process itself and minimise other variables
affecting overall RNAi efficacy, we used a robust co-transfection system to
quantitatively assess the relative potencies of 21 small-hairpin (sh) RNA
vectors, targeting conserved sites throughout the HBV genome, against viral
RNAs, proteins, nucleocapsids, and secreted virions under standardised
conditions.
Results
The approach enabled a distinct efficacy ranking, with the six most potent
shRNAs achieving 95% reductions in virion formation, sequence-specifically and
without detectable interferon induction, yet by differentially affecting
different Background & Aims
RNA interference (RNAi) may offer new treatment options for chronic hepatitis
B. Replicating via an RNA intermediate, hepatitis B virus (HBV) is known to be
principally vulnerable to RNAi. However, beyond delivery, the relevant issues
of potential off-target effects, target site conservation in circulating HBV
strains, and efficacy of RNAi itself have not systematically been addressed,
nor can the different existing data be quantitatively compared. The aim of
this study was to provide such information.
Methods
To focus on the intracellular RNAi process itself and minimise other variables
affecting overall RNAi efficacy, we used a robust co-transfection system to
quantitatively assess the relative potencies of 21 small-hairpin (sh) RNA
vectors, targeting conserved sites throughout the HBV genome, against viral
RNAs, proteins, nucleocapsids, and secreted virions under standardised
conditions.
Results
The approach enabled a distinct efficacy ranking, with the six most potent
shRNAs achieving 95% reductions in virion formation, sequence-specifically and
without detectable interferon induction, yet by differentially affecting
different steps. Efficacy correlated poorly with predictions and was not
principally abolished by target structure. Sequence comparisons suggest that
truly conserved, RNAi-targetable sequences comprise less than 500 nucleotides
of the circulating HBV genomes.

steps. Efficacy correlated poorly with predictions and was not
principally abolished by target structure. Sequence comparisons suggest that
truly conserved, RNAi-targetable sequences comprise less than 500 nucleotides
of the circulating HBV genomes.

咬牙硬挺

什么意思啊？只要大意就好！

StephenW

J Hepatol. 2010 Jun;52(6):817-26. Epub  2010 Apr 1.
Quantitative assessment of the antiviral potencies of 21 shRNA vectors targeting conserved, including structured, hepatitis B virus sites.Sun D, Rösler C, Kidd-Ljunggren K, Nassal M.
SourceBethune International Peace Hospital, Departmrnt of Liver Disease, 398 West Zhongshan Road, 050082 Shijiazhuang, PR China.

AbstractBACKGROUND & AIMS: RNA interference (RNAi) may offer new treatment options for chronic hepatitis B. Replicating via an RNA intermediate, hepatitis B virus (HBV) is known to be principally vulnerable to RNAi. However, beyond delivery, the relevant issues of potential off-target effects, target site conservation in circulating HBV strains, and efficacy of RNAi itself have not systematically been addressed, nor can the different existing data be quantitatively compared. The aim of this study was to provide such information.
METHODS: To focus on the intracellular RNAi process itself and minimise other variables affecting overall RNAi efficacy, we used a robust co-transfection system to quantitatively assess the relative potencies of 21 small-hairpin (sh) RNA vectors, targeting conserved sites throughout the HBV genome, against viral RNAs, proteins, nucleocapsids, and secreted virions under standardised conditions.
RESULTS: The approach enabled a distinct efficacy ranking, with the six most potent shRNAs achieving 95% reductions in virion formation, sequence-specifically and without detectable interferon induction, yet by differentially affecting different steps. Efficacy correlated poorly with predictions and was not principally abolished by target structure. Sequence comparisons suggest that truly conserved, RNAi-targetable sequences comprise less than 500 nucleotides of the circulating HBV genomes.
CONCLUSIONS: The HBV genome can harbour only a finite number of optimal target sites, but current predictions are poorly suited to constrain the number of possible candidates. However, the small size of the highly conserved sequence space suggests experimental identification as a viable option.
Copyright 2010 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

StephenW

J肝胆病。 2010年6月，52（6）:817 - 26。作者2010年4月1日。
21 shRNA的目标保守，包括结构，乙肝病毒的网站载体的抗病毒药物的效力的定量评估。
罗斯勒，基德，Ljunggren Nassal M · K，孙一丁，
来源

，中国公关，050082石家庄白求恩国际和平医院，肝病Departmrnt，中山西路398。
摘要
背景及目的：

RNA干扰（RNAi）可以提供新的治疗慢性乙型肝炎的选项，通过RNA中间体，乙肝病毒（HBV）是已知的主要脆弱的RNAi的复制。然而，超出交付，潜在的脱靶效应，循环乙肝病毒株的目标遗址保护，和RNAi技术本身的疗效有关的问题还没有系统地解决，也可以定量比较不同的现有数据。本研究的目的是提供此类信息。
方法：

要集中在细胞内的RNAi过程本身，并减少其他变量影响RNAi的整体功效，我们用一个强大的共转染系统定量评估相对的效力21（SH）小发夹RNA的载体，针对整个HBV基因组的保守网站，对病毒的RNA，蛋白质，核衣壳，和标准化的条件下分泌的病毒颗粒。
结果：

方法使实现六个最有力的shRNA在病毒粒子形成的95％减少一个独特的疗效排名，具体，没有检测到干扰素诱导序列，但差异影响不同的步骤。疗效不佳相关的预测和主要目标结构并没有取消。序列比较表明，真正的保守，RNAi技术可定位的序列包括不少于500循环的乙肝病毒基因组核苷酸。
结论：

HBV基因组的海港只有一个最佳目标网站的数量有限，但不适合目前的预测是可能的候选人数限制。然而，高度保守的序列空间的体积小，表明实验鉴定作为一种可行的选择。

2010年欧洲肝脏研究协会。发布由Elsevier B.五，保留所有权利。