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Publication : Efficient poly A trap approach allows the capture of genes specifically active in differentiated embryonic stem cells and in mouse embryos.

First Author  Salminen M Year  1998
Journal  Dev Dyn Volume  212
Issue  2 Pages  326-33
PubMed ID  9626507 Mgi Jnum  J:164347
Mgi Id  MGI:4833664 Doi  10.1002/(SICI)1097-0177(199806)212:2<326::AID-AJA17>3.0.CO;2-1
Citation  Salminen M, et al. (1998) Efficient poly A trap approach allows the capture of genes specifically active in differentiated embryonic stem cells and in mouse embryos. Dev Dyn 212(2):326-33
abstractText  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 beta-galactosidase (beta-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 beta-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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