OmicsLink™ shRNA表达克隆

产品应用

基因表达下调 — 以包含无义序列的克隆为对照，研究shRNA克隆对单个基因表达的下调效果。在每个shRNA克隆订单中赠送对照克隆。

信号通路研究 — 我们将克隆按照不同的信号传导、代谢、疾病通路、基因家族等进行了分类，并将已知同一通路的shRNA表达克隆以96孔或384孔板的形式排列在一起，可一次研究一组基因在某一信号通路中的功能。目前我们提供针对人类全部激酶基因的shRNA表达克隆，并已按照按不同激酶的功能进行了分类。

靶点及功能验证 — OmicsLink™ ORF表达克隆和在shRNA靶序列带有沉默突变的ORF表达克隆一起使用可用于shRNA靶点验证和基因功能拯救研究。

图2. shRNA抑制基因表达的机制

qRT-PCR — shRNA克隆的表达能导致靶标基因所转录的mRNA被切割和降解，从而降低蛋白的表达量。所以推荐检测表达抑制效率的第一步是qRT-PCR。GeneCopeia提供验证过的对应靶标基因和其他已报导的转录本的qRT-PCR引物。

Western Blot检测 — 大部分情况下，检测shRNA对基因表达的抑制效率需要用Western Blot方法来检测蛋白表达水平。

功能检测 — 检测表达抑制效率还可用生物和生化的终点分析方法，如细胞增殖、克隆形成、细胞周期、细胞迁移等。

报告基因检测表达水平 — 当研究的靶标基因内源转录水平很低，且不适宜使用qRT-PCR和WB检测时，表达抑制效率的检测也可通过共转导shRNA克隆和一个含靶标基因并带了报告基因的表达克隆，检测报告基因的表达水平即可。靶基因的mRNA遭shRNA的破坏，同时导致了报告基因的mRNA遭破坏，从而表现为报告蛋白的表达量下降。因而表达抑制效率可以用报告蛋白的酶活性分析结果来进行量化。

图3. shRNA(psi-sH1载体)对人激酶家族成员表达抑制效率的验证。psiCheck-AP载体以CMV为启动子，包括靶标激酶的ORF区和用于比色实验的碱性磷酸酶（AP）基因。A. 通过比色实验量化AP在没有shRNA的情况下的表达。B. 共转染shRNA和AP ORF表达克隆后，AP ORF的转录本被降解，AP基因翻译受阻导致荧光消失。

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OmicsLink shRNA表达克隆载体  

 shRNA表达克隆客户定制服务

预制的shRNA克隆套装

GeneCopeia提供载体可选的超过24,000条人、20,000条小鼠和10,000条大鼠预制的shRNA克隆套装。

慢病毒载体的shRNA表达克隆

GeneCopoeia 提供定制慢病毒载体的shRNA表达克隆的服务， 浏览价格信息，请填写表格。

shRNA表达克隆定制服务

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