Gene overexpression - Targeted insertion : knock-in mouse |
|
This approach is based on targeted insertion of the transgene in a selected locus. Using homologous recombination in Embryonic Stem cells (ES cell), it is possible to insert the transgene in an exact position. This approach is called "knock-in" (knockin). |
|
| Advantages |
Drawbacks |
Specific integration site
Transgene genetic environment is totally controlled
Only one transgenic line is necessary
|
Molecular biology work can be more complex and longer
|
|
|
|
Intellectual Property
Knock-in is a general term for models expressing a transgene targeted in a
selected locus. Several technologies exist to create such models. These
technologies use techniques (alone or in combination) such as homologous
recombination, recombinases, insertion shuttles, etc.
genOway can implement technologies that are free of use for customers and in
particular that are not depending from the Pasteur Institute patent family
(abusively called Homologous recombination technology by Cellectis).
|
|
The offer genOway has developed can include the following steps, but the service is flexible (can be limited to part of this offer and/or adapted to customer requirements) : |
|
| Step 1 : |
Subcloning and characterization of the locus of interest |
| Step 2 : |
Cloning and characterization of the cDNA of interest |
| Step 3 : |
Knock-in / Knockin vector construction (targeting vector) |
| Step 4 : |
Homologous recombination in Embryonic Stem cells (ES Cell) |
| Step 5 : |
Blastocyst injection and chimera generation |
| Step 6 : |
Breeding to the F1 and F2 generation |
|
|
The "knock-in" (knockin) model provides a very reliable way of overexpressing a transgene. Technological solutions have been developed and fully validated at genOway to shorten the development time of such models. Please visit our " Quick Knock-in™" page |
|
The development of the first cloned rat reported by genOway in Science ( Zhou et al. 2003) paves the way for the development of overexpression rat models based on targeted insertions. This knock in approach will provide researchers with rat models of high biological quality. Even though this technology is not yet available as a service, research partnerships may be established. |
 | |
|
|
Service offer
Ready-to-use models