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Efficient poly A trap approach allows the capture of genes specifically active in differentiated embryonic stem cells and in mouse embryos by Marjo Salminen; Barbara I. Meyer; Peter Gruss is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Special vectors have been constructed that allow the trapping of genes in mouse embryonic stem (ES) cells. These vectors generally contain the neomycin phosphotransferase (neo) gene for selection and the ␤ -galactosidase (␤-gal) gene as a marker. Promoterless vectors can be used to identify genes that are active in undifferentiated ES cells. To also have access to genes that are inactive in totipotent ES cells, we constructed a polyadenylation (poly A) trap vector in which the expression of a poly A less neo gene is driven by a constitutive promoter, whereas the expression of ␤-gal depends on the trapped sequences. We demonstrate here that this vector integrates with a high frequency into transcription units and that it traps genes with very different expression patterns in vitro and in vivo. The vector integrates efficiently into transcription units that are inactive in undifferentiated ES cells and which can be activated through in vitro differentiation. Furthermore, in vivo expression patterns demonstrate that this vector integrates into genes that exhibit a highly specific temporal and spatial expression pattern during embryogenesis.

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Author

Marjo Salminen; Barbara I. Meyer; Peter Gruss

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 1058-8388)

Published

1998

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)