基本信息
別稱: psiCHECK2
啟動子: SV40
復制子: pUC
終止子: SV40 poly(A) signal
質(zhì)粒分類: 哺乳細胞，信號通路報告載體
質(zhì)粒大小: 6273bp
原核抗性: Amp
篩選標記: Luc/hRluc
克隆菌株: DH5a
培養(yǎng)條件: 37 度
表達宿主: 哺乳細胞
誘導方式: 無須誘導，瞬時表達
5'測序引物: psiCHECK-2-F：GAAGTTCCCTAACACCGAGT
3'測序引物: psiCHECK-2-R：CCCCCGCGAGGTCCGAAGAC
質(zhì)粒屬性
載體宿主： 哺乳細胞
載體用途： 信號報告
基因種屬： 空載體
基因類型： shRNA,miRNA,UTR
原核抗性： Amp
熒光蛋白： fLuc,rLuc
質(zhì)粒簡介
psiCHECK-2 Vector is designed to provide a quantitative and rapid approach for optimization of RNA interference (RNAi). The vectors enable the monitoring of changes in expression of a target gene fused to the reporter gene. In both vectors, Renilla luciferase is used as a primary reporter gene, and the gene of interest can be cloned into the multiple cloning region located downstream of the Renilla luciferase translational stop codon. Initiation of the RNAi process toward a gene of interest results in cleavage and subsequent degradation of fusion mRNA. Measurement of decreased Renilla luciferase activity is a convenient indicator of RNAi effect .
RNAi is a phenomenon by which double-stranded RNA complementary to a target mRNA can specifically inactivate gene function by stimulating the degradation of the target mRNA . Because of the ability to inactivate genes, RNAi has emerged as a powerful tool for analyzing gene function.
In mammalian systems, including cultured mammalian cells, chemically synthesized double-stranded short interfering RNA molecules (<30 nucleotides; siRNA) result in dsRNA duplexes <30 base pairs in length that induce RNAi .RNAi duplexes >30bp induce the interferon response and nonspecific degradation of mRNA and cannot be used as tools for specific gene silencing .
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