Rosa26 Targeting Overexpression

This technology relies on transgene integration in the targeted Rosa26 locus to overcome classical random transgenesis drawbacks.

This is achieved through homologous recombination in ES cells of a targeting vector containing the transgene (conditional in most of the cases) and Rosa26 homology arms. Alternative docking sites are Hprt or Col1a1.

Advantages

• Transgene insertion in a specific locus
• Integration of a single-copy transgene; this minimizes the risk of silencing by methylation as the Rosa26 has been used extensively for transgene expression.
• Transgene consistent levels of ubiquitous expression driven either by the endogenous ROSA26 promoter in the embryo or by a synthetic CMV enhancer-chicken b-actin promoter (pCAG) in later stages; all founder lines will exhibit similar expression level

Drawbacks

• This method cannot be used to generate knock-out or knock-in models

PHENOMIN expertise

• Personalized genome engineering service to fit our customer needs
• Implication in international consortia (near 20 different consortia) to benefit from the last technics and scientific expertise
• Continuous R&D activity to improve our technics and success rate and propose our customers the best approach for their projects

Publications

• Birling et al., 2012, Highly-efficient, fluorescent, locus directed Cre and FlpO delete mice on a pure C57BL/6N genetic background