products | Mirus

Mirus Corp. - Gene Transfer Specialists

For more information please visit the
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or download the Mirus catalog (1.8 MB pdf).
 

Below you will find some selected Mirus products.

siXpress^TM PCR Vector Systems

• Enables efficient in situ expression of siRNA and knockdown.
  
• Synthesize multiple siRNA expression cassettes by PCR in hours.
  
• Select the siRNA expression cassette that provides the most desirable effect.
  
• Option of cloning PCR generated expression cassette in supplied vector.
  
• Complete Systems: provided with primers, template, controls and TransIT®-LT1 Transfection Reagent.
  

The siXpress™ PCR Vector Systems contain reagents required for the optimized preparation and transfection of siRNA expression cassettes. Mirus offers three siXpress™ PCR Vector Systems, each utilizing a different polymerase III promoter: human U6, mouse U6, and human H1 (individual cell lines may vary in their promoter preferences). Double-stranded DNA containing the siRNA expression cassette is generated by polymerase chain reaction (PCR) and transfected into mammalian cells in vitro using TransIT®-LT1 Transfection Reagent. Following nuclear transcription of the siRNA expression cassette, siRNA transcripts are exported into the cytoplasm and enter the RNAi pathway. RNAi applications include verification of effective siRNA sequences, studies of gene function, drug development, target validation, and other biological studies. The optimal PCR-generated expression cassettes can also be cloned into the supplied Template/Cloning Vector and grown under kanamycin selection in bacteria to provide a large amount of plasmid for further gene silencing experiments. Each siXpress™ PCR Vector System provides sufficient materials to generate 20 siRNA expression cassettes. The supplied Luciferase Control Vector generates an siRNA hairpin directed against the firefly luciferase (luc+) gene (Promega). PCR products generated from this vector can be used as negative controls in experiments with user-designed siRNA expression cassettes. The Control Vector can also be used as a positive control in cells transiently or stably expressing luciferase.

CHO-luc cells (CHO-K1 cells expressing a stably integrated firefly luciferase), were transfected with siXpress™ PCR siRNA expression cassettes using TransIT®-LT1 Transfection Reagent. Three siXpress™ promoter vectors were tested: human U6, mouse U6, and human H1. An untargeted SEAP (secreted alkaline phosphatase) hairpin siRNA expression cassette (blue), a luciferase specific hairpin siRNA expression cassette (orange), and a luciferase specific hairpin siRNA control cassette (yellow) were amplified for each promoter. The luciferase specific hairpin siRNA control cassette was amplified directly from the corresponding luciferase hairpin siRNA expression plasmid. 500 ng of each cassette were transfected with TransIT®-LT1 Transfection Reagent and assayed after 48 hours. Luciferase siXpress™ cassettes and luciferase control cassettes were normalized to the SEAP cassettes for each corresponding promoter.

MiraCLEAN^® Endotoxin Removal Kit

Features

• Efficient removal of endotoxin from nucleic acid preparations.
  
• Ideal for in vivo and in vitro applications.
  
• Improved yields via distinct visualization of extraction phases.
  
• Simple protocol.
  

Many molecular biology laboratory applications require endotoxin-free preparations of plasmid DNA and high molecular weight genomic DNA. The MiraCLEAN^® Endotoxin Removal Kit provides a convenient and improved method for the removal of bacterial endotoxins from nucleic acids for both in vivo and in vitro applications. Esherichia coli, a Gram-negative eubacteria, is the common host for plasmid production. The outer leaflet of the outer membrane contains lipopolysaccharides (LPS, or endotoxin) which can cause inflammatory reactions, fever and endotoxic shock in vivo. Endotoxin in plasmid preparations is also known to decrease transfection efficiencies in vitro. The MiraCLEAN^® Kit is based on a rapid phase extraction of endotoxin. The proprietary pink-colored EndoGO Extraction Reagent allows better visualization of the interface between phases, thereby greatly facilitating phase separation and increasing recovery of nucleic acid. EndoGO Extraction Reagent and the MiraCLEAN^® Buffer remove endotoxin contamination from DNA and thus, aid in the safety and efficiency of gene delivery research.

The data show the average luciferase expression resulting from duplicate transfections completed on COS-7 cells in 35 mm plates in complete growth media. Cells were grown to approximately 60% confluency and transfected using 2 µg plasmid DNA (pCI-Luc)/2µl of TransIT®-LT1 Reagent. Cells were harvested 24 hours post transfection and assayed for luciferase expression.

More about TransIT Transfection Reagents ...

Download: MiraCLEAN Endotoxin Removal Kit product information

pLIVE^TM In Vivo Expression and Reporter Vectors

Features

• Designed for high level, prolonged expression of transgenes in the mouse liver.
  
• Long term liver expression through 8 months.
  
• Available with positive control vectors expressing either lacZ or SEAP.
  
• Compatible with hydrodynamic delivery using the TransIT^®-EE and TransIT^®-QR Hydrodynamic Delivery Solutions and other liver delivery methodologies.
  

As a leader in the development of the hydrodynamic tail vein injection technique for the delivery of nucleic acids to the mouse liver, Mirus Bio Corporation has developed a state of the art liver-specific transgene expression vector, the pLIVE™ (Liver In Vivo Expression) Vector. The pLIVE™ Vector is designed for high level, prolonged expression of transgenes in mouse liver. This vector utilizes a chimeric promoter composed of the minimal mouse albumin promoter and the mouse alpha fetoprotein enhancer II. Two introns have been engineered into the expression plasmid so that they will be present in the primary transcript, in turn increasing expression of the transgene. Downstream of the first intron is a multiple cloning site (MCS) with eight unique restriction sites allowing for simple insertion of the gene of interest. Together this chimeric promoter and the two introns are capable of promoting high level transgene expression in the liver for extended lengths of time compared to classic promoters such as the cytomegalovirus immediate early promoter (CMV).
In addition to the pLIVE™ Vector, two reporter vectors derived from the pLIVE™ Vector, the pLIVE™-lacZ and pLIVE™-SEAP Vectors, can be used as positive controls. Expression of the E. coli lacZ gene from pLIVE™-lacZ Vector can be monitored in the liver using either classical X-gal staining of liver tissue sections or quantitative ß-galactosidase assays using liver lysates. Expression of human placental secreted alkaline phosphatase (SEAP) from the pLIVE™-SEAP Vector can be easily monitored using a quantitative assay of mouse serum following delivery. The high level, long term liver-specific expression of transgenes from the pLIVE™ Expression Vector, as well as the availability of the positive control pLIVE™-lacZ and pLIVE™-SEAP Reporter Vectors, make the pLIVE™ Vector series the ideal choice for in vivo liver expression studies in mice.

pLIVE™ Vector: download map (pdf 279 KB)

pLIVE™-lacZ Vector : download map (pdf 282 KB)

pLIVE™-SEAP Vector: downlaod map (pdf 267 KB)

Please download: pLIVE product information (pdf 136 KB)