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Transgene service of the Tol2 system

1. Brief introduction of the technology
The Tol2 transposon was first found in the fish body, so it is widely used in the study of zebrafish, and has high transgene efficiency. It has high transposition efficiency in the vertebrates besides zebrafish, and application in human and mouse cells has been widely reported. The transposition efficiency of Tol2 is higher in comparison to the commonly-used transposons Sleeping Beauty and PiggyBac. Several cell lines and animal experiments show that the transgene mediated by the Tol2 system has no obvious specificity for insertion sites. The inserted section’s size will not influence the normal operation of the Tol2 system. BIOCYTOGEN used the Tol2 system place a transgene on the 13kb section of the mouse, and obtained a higher efficiency. We used this system to successfully construct transgenic rats and stable cell lines. Therefore, the Tol2 system is a powerful tool for construction of transgenic animal models and cell lines.


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2. Advantages of Tol2 transgene system
• Higher transgene insertion efficiency
• No obvious insertion site bias
• Can insert large exogenous DNA—a 13kb section has been transferred in successfully

3. Application of Tol2 transgene system
3.1 Construct the gene expression cell line simply and with high efficiency
The Tol2 system is simple compared with establishment of a stable cell strain with plasmid transfection and drug resistance screening, and has a high success rate and requires only a short period of time. the Tol2 system is easy to operate and is low cost compared with the virus system,.

3.2 Preparation of transgenic rat and mouse
In comparison to the conventional DNA pronuclear injection, the Tol2 system instead uses cytoplasmic injection, which is easy to do and has a high success rate. This method overcomes the low success rate of transgenic rat prepation via conventional DNA pronuclear injection.

4. Service type

Service type

Quality control method

Product form

Transgenic cell line

PCR genotype detection with two pairs of primers

Frozen cell

Transgenic mouse

PCR genotype detection with two pairs of primers

Mouse of generation F0

Transgenic rat

PCR genotype detection with two pairs of primers

Rat of generation F0

 

5. Difference comparison of the different methods to prepare the transgenic cell lines

Preparation method

Efficiency

Preparation method

Stability

DNA direct transfection

Low (~5%), and long period

Simple, and transfect the DNA directly

Unstable

Tol2 transposons method

High (~20%), and short period

Simple, one-time cloning,  co-transfection with the transposase

Stable

Retrovirus-mediated gene transfer

High (~100%)

Preparation is complex, and virus packaging and separation is needed

Stable


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