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Publication : An efficient gene-trap method using poly A trap vectors and characterization of gene-trap events.

First Author  Niwa H Year  1993
Journal  J Biochem Volume  113
Issue  3 Pages  343-9
PubMed ID  8387481 Mgi Jnum  J:40067
Mgi Id  MGI:87406 Doi  10.1093/oxfordjournals.jbchem.a124049
Citation  Niwa H, et al. (1993) An efficient gene-trap method using poly A trap vectors and characterization of gene-trap events. J Biochem 113(3):343-9
abstractText  New trap vectors (U1 and U2) have been developed to trap genes in murine embryonic stem (ES) cells. The polyA addition signal of the neomycin phosphotransferase II (neo) gene was removed from these vectors so that they needed to trap an endogenous polyA signal for expression of the neo gene. The frequency of gene-trap events of these vectors was about five times higher than with the vector containing the polyA signal, and only one copy of the trap vector was integrated in most cases. Four out of five 5'-flanking regions of the integrated vector in ES cell lines were found to be novel endogenous promoters, suggesting that this method is efficient for trapping genes in ES cells. In two cases analyzed, large deletions or rearrangements spanning more than 10 kb were found in the 3'-flanking region of the trap vector introduced by electroporation. This result suggests that phenotypes observed in homozygotes with a mutated allele could be due to the disruption of a gene adjacent to the trapped gene, but not of the trapped gene.
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