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RNA Silencing Mechanism
- dsRNA is first degraded by Dicer (a highly conserved RNase) into siRNA (~22 bp).
- siRNA binds to RISC (RNA Induced Silencing Complex) to form siRNA-Protein Complex.
- The Complex cleaves mRNA with a complementary sequence, inhibiting downstream expression.
A Successful RNA Silencing Expriment
Transfection Methods
The common methods of mammalian transfection include:
- calcium phosphate coprecipitation
- electroporation
- microinjection
- gene gun
- liposome transfection
- etc
Currently, liposome transfection is most commonly used.
Notes for Liposome Transfection
Several aspects need to be paid attention to for liposome transfection:
- the type and dosage of transfection reagent
- the type and dosage of siRNA
- the cell density
- the operation sequence
- the water bath timing of cells and siRNA-transfection reagent complex
Chemically Modified siRNA
The most difficult problem of RNAi technology is the stability of siRNA. Here, with chemical modification, our chemically modified siRNA has much higher stability than the common siRNA. The chemical modification not only enhances the life span of siRNA in serum and cell culture but also enhances its activity in vitro.
Sirnafectamine Transfection Reagent
Sirnafectamine is a new generation of the RNA transfection reagent, which can be used in RNA transfection experiments. Sirnafectamine accelerates the binding of glycoprotein on cell membrane, which promotes the endocytosis of cells. In addition, compared with other siRNA transfection reagents, Sirnafectamine has minimal cytotoxicity and the best cell state after transfection.
SBS Genetech is a long-term sponsor of
Cold Spring Harbor Laboratory
