| Infection of Human Hepatocyte-chimeric Mice with HBV and in vivo Treatment with εRNA
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Author:
Date:
2016-01-20
[Abstract] Hepatitis B virus (HBV) can cause both acute and chronic disease in human liver with potentially high risk of cirrhosis and liver cancer. The host range of non-human primates susceptible to this virus is limited. Therefore, experimental studies with human hepatocyte-chimeric mice provide an invaluable source of information regarding the biology and pathogenesis of HBV. This section describes the protocol for infection of the human hepatocyte-chimeric mice with HBV. In addition, it has recently been shown that HBV replication can be suppressed by exogenous expression of viral epsilon RNA (εRNA; Sato et al., 2015), which serves as an encapsidation signal (Bartenschlager et al., 1992). Based upon this finding, we also describe the protocol for the liposome-mediated delivery ...
[摘要] 乙型肝炎病毒（HBV）可引起人类肝脏的急性和慢性疾病，具有潜在的肝硬化和肝癌的高风险。 对这种病毒敏感的非人灵长类动物的宿主范围有限。 因此，用人肝细胞嵌合小鼠的实验研究提供了关于HBV的生物学和发病机理的非常宝贵的信息来源。 本节描述了用HBV感染人肝细胞嵌合小鼠的方案。 此外，最近已经表明，HBV复制可以通过作为壳体化信号的病毒ε-RNA（εRNA; Sato等人，2015）的外源表达来抑制（Bartenschlager et al。，1992）。 基于这一发现，我们还描述了脂质体介导的在这些嵌合小鼠中将编码εRNA的质粒递送至肝的方案。
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| Single-cell Gene Expression Profiling of Mouse Stem Cells With Fluidigm BiomarkTM Dynamic Array
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Author:
Date:
2013-05-05
[Abstract] This protocol describes how to use Fluidigm BiomarkTM 96.96 dynamic arrays for high-throughput expression profiling from single mouse stem cells, assaying up to 96 independent samples with up to 96 quantitative PCR (qPCR) probes (equivalent to 9,216 reactions) in a single experiment. This Dynamic Array contains a network of microfluidic channels, chambers and valves that automatically assemble all these individual PCR reactions. Single-cell profiling can provide definitive evidence of stem cell heterogeneity. Modifications are most likely needed if users intend to use BiomarkTM 48.48 Dynamic array or experimenter-designed primers in conjunction with DNA-binding dyes such as EvaGreen (Biotium 31000).
[摘要] 该方案描述了如何使用Fluidigm Biomark？sup？9696动态阵列从单个小鼠干细胞进行高通量表达谱分析，使用多达96个定量PCR（qPCR）探针测定多达96个独立样品（相当于 9,216个反应）。 该动态阵列包含微流体通道，腔室和阀门的网络，其自动组装所有这些单独的PCR反应。 单细胞谱分析可以提供干细胞异质性的明确证据。 如果用户打算使用Biomark TM 48.48动态阵列或实验设计的引物与DNA结合染料如EvaGreen（Biotium 31000）结合，则很可能需要进行修饰。
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