Transfection reagent for short oligonucleotides

LipoTrust™ EX Oligo


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Application

Transfection reagent for short oligonucleotides such as siRNA, antisense DNA or miRNA.

Feature and Advantages

• Cationic liposome formulation
• Highly efficient delivery of short oligonucleotides in the presence of serum (please see blow)
• Simple handling: direct mixing of short oligonucleotide into lyophilized powder

Improvement of siRNA stability in the serum mixed with liposome
SiRNA easily decomposes in a few minutes by Nuclease in the presence of serum whereas siRNA mixed with appropriate liposome can survive for a few hours. In fact, more than 50% of siRNA activity remained in 120 minutes with LipoTrust™ EX Oligo while 99 % of naked siRNA decomposed less than 5 minutes in the presence of bovine serum.
Both free siRNA and siRNA encapsulated in liposome were allowed to incubate in the bovine serum for more than 2 hours. Aliquots of serums were taken intermittently and respective RNAs were extracted .followed by siRNA contents analysis with HPLC.
As shown in the left Figure, free siRNA decomposed rapidly while as much as 50% of siRNA encapsulated in liposome were found to survive during 120 minutes incubation.

Kit component

• Lipo Trust™ EX Oligo (Lyophilizate for 1ml) x1
• Instruction manual

Example for in vivo use

Reduction of both mice serum cholesterol and LDL by injecting LipoTrust™ EX Oligo / siRNA complex which can down-regulate Apo-B gene.

Procedure
1. 1 ml of prepared 83.0 pmol/μL of siRNA solution which can down-regulate Apo-B gene was poured into LipoTrust™ EX Oligo vial. Liquid was mixed well and kept for 20 minutes according to the Protocol.
2. Aliquot of each 80 μl were injected into 6 week old mice (n=5) from tail vein once in a day.
3. The same volume of injection was continuously repeated for 3 days and 80 μl of blood were taken 48 hours after final injection.
4. Blood was allowed for ultra-centrifugation with 10,000 rpm for 10 minutes.
5. Serum was taken, and then both cholesterol and LDL were analyzed.
6. For the comparison, 1 mL of LipoTrust™ EX Oligo liposome without containing siRNA was prepared and injected into mice (n=5) with the same volume and same interval.

Result
As shown in the right figure, more than 10% reduction of both cholesterol and LDL using siRNA mixed liposome were observed in comparison with that of siRNA-free liposome.

Product List

Product Name Cat# Quantity Price

LipoTrustTM EX Oligo for in vitro DataSheet ,

CSR-LEO-01-EX

1 ML ¥46,200
$616

LipoTrustTM EX Oligo for in vivo DataSheet ,

CSR-LEO-10-EX

1 ML ¥144,000
$1920

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.



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