The highly effective service to establish stable cell lines is now available at Creative BioMart. We adopt the Tol2 transposon to integrate the GOI into the genome of the host cell permanently to establish the stable protein expression cell line. The establishment process is mediated by the non-virus vector and according to our experience, the system enables sequence insertion up to 11-13kb.
The technology is initially developed from Tol2 transposon which was isolated from teleost fish. Tol2 is a class II transposon. The Tol2 system typically consists of two vectors:
A.Helper vector: encodes the transposase which mediates the sequence transposition.
B.Transposon plasmid: contains the GOI as well as two inverted terminal repeats (ITRs)
When co-transfecting the helper vector and transposon plasmid into the host cell, the transposase expressed by the helper vector will recognize the ITR and cut it from the transposon vector and insert it into host cell chromosome. (It should be noted that the insertion site of Tol2 transposon in host cells has no obvious base preference.)
As the efficiency of co-transfection may affect the integration efficiency, if required, we can also transcribe Tol2 mRNA in vitro and inject them into host cells. Just let us know your requirement.
With the loss of helper vector or the degradation of transposase mRNA, GOI with two ITRs are permanently integrated into the host genome.
A.Optimized Vector
The helper vector and the transposon plasmid had been reformed by Creative BioMart’s technical group for high copy number replication in E. coli. The quality test shows that our Tol 2 system is highly efficient for a wide range of mammalian cells. The expression level is guaranteed.
B.High Capacity
The vector can carry a large fragment of foreign DNA, up to 13kb.
C. No significant bias of insertion site
D. Technical simplicity
There is no need for virus packaging. Thus, it is far easier to be operated than those viral vector-mediated systems.
Here are some recommended applications of our Tol2 Vector-Mediated Stable Cell Line Establish Services.
A.Simple and efficient establishment of stable gene expression cell lines
Compared with plasmid transfection and drug resistance screening to establish stable expression cell lines, Tol2 system is simple, with high success rate and short cycle. Compared with the viral plasmid-mediated system, the experiment operation is simple and the cost is low.
B.Preparation of transgenic rats and mice (Tol2 homology)
Compared with the traditional DNA prokaryotic injection, Tol2 system adopts cytoplasmic injection, which is easy to operate and has high success rate. In particular, it overcomes the problem of low success rate of traditional DNA prokaryotic injection preparation of transgenic rats.
Creative BioMart is always devoted to providing high-quality and satisfactory service to our customers. If you are interested in our services or have any questions, please feel free to contact us or make an online inquiry.
