|Products||pMIR-hU6-Luc, Human U6 Luciferase Control Vector from Mirus Bio Corporation|
|Company||Mirus Bio Corporation|
|Item||pMIR-hU6-Luc, Human U6 Luciferase Control Vector|
|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. 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. 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, in a plasmid [2,3,5-8], in a viral vector, or integrated into the genome. 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. 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).
PCR Expression Cassettes can be cloned using either the Hind III or EcoR I site upstream of the promoter and Xho I or BamH I site downstream of the siRNA and termination sequence. The cloning sites used must be unique in the PCR product. Hind III and Xho I are recommended because these restriction enzymes function with compatible buffers.
|Info|| Mirus Bio Corporation|
505 S. Rosa Rd.
Madison, WI 53719
Fax Number: (608) 441-2849
Web Site: http://www.mirusbio.com