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Publication : Co-expression of multiple transgenes in mouse CNS: a comparison of strategies.

First Author  Jankowsky JL Year  2001
Journal  Biomol Eng Volume  17
Issue  6 Pages  157-65
PubMed ID  11337275 Mgi Jnum  J:78664
Mgi Id  MGI:2385666 Doi  10.1016/s1389-0344(01)00067-3
Citation  Jankowsky JL, et al. (2001) Co-expression of multiple transgenes in mouse CNS: a comparison of strategies. Biomol Eng 17(6):157-65
abstractText  The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.
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