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pMIR-mU6, Mouse U6Template (Vector) from Mirus Bio Corporation

ProductspMIR-mU6, Mouse U6Template (Vector) from Mirus Bio Corporation
Company Mirus Bio Corporation
Item pMIR-mU6, Mouse U6Template (Vector)
Price $147.00
Description The potential applications for the use of small interfering RNA (siRNA) molecules to silence gene expression via the RNA interference (RNAi) pathway have generated tremendous interest in the cellular and molecular biology fields. siRNA for experimental use can be synthesized in vitro, either chemically or enzymatically, and then transfected into the target cells in vitro or in vivo [1-4]. A convenient alternative to introduce siRNAs into mammalian cells is transient or stable transfection of expression constructs that drive siRNA transcription in the nucleus from RNA Polymerase III promoters [2,3,5-11]. An expression cassette is a DNA construct that includes a complete transcription unit consisting of a promoter, a template sequence to be transcribed by RNA polymerase, and a transcription termination signal. An siRNA expression cassette can be in a linear fragment of DNA [12], in a plasmid [2,3,5-8], in a viral vector [9-11], or integrated into the genome [13]. The expression cassettes that have been used for generating siRNA in mammalian cells have utilized the human U6, human H1, or mouse U6 RNA polymerase III promoters. These promoters drive expression of short RNA transcripts that can become siRNA.

An siRNA molecule is formed from short complementary sense and anti-sense strands of RNA. The sense strand of the siRNA has the same sequence as a short region of the mRNA targeted for RNAi. Each siRNA strand may originate from different transcripts expressed in the same cell or both siRNA strands may be part of a single transcript that folds back to form a short hairpin structure [2] (Figure 1). The expression cassette(s) encoding the siRNA strands can be delivered to cells in the same fragment of DNA (hairpin expression cassette) or on separate fragments of DNA (sense and anti-sense strand expression cassettes; Figures 1 and 2).

Although PCR based siRNA expression cassettes have the advantages of being fast to produce and test, some users may choose to transfect the siRNA expression cassette in the form of a plasmid or to have a large supply of expression cassette DNA readily available. Any of the PCR products generated in the previous sections can be cloned into the siXpress(TM) Template if the target-specific downstream primer used to generate the PCR products include an Xho I or BamH I restriction site. For additional instruction on molecular biology procedures, refer to Sambrook et al.

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