Evidence for the stochastic integration of gene trap vectors into the mouse germline.
A large scale insertional mutagenesis experiment was performed in embryonic stem (ES) cells by introducing two types of gene trap vectors into the genome. These cell lines carrying mutations were introduced into the mouse germline. In order to assess the feasibility of a large scale cloning of the targeted genes from these lines, we have isolated and characterized 55 trapped exons from the corresponding ES cells. Analysis of the data has revealed that vectors containing or lacking an internal ribosome entry site (IRES) can integrate into the ES cell genome stochastically. The targeted genes comprise 30% known genes, 20% expressed sequence tags (ESTs) and 50% novel or unknown genes. The known genes belong to several major classes and represent complete or partial knockouts. Using currently available methods or modifications of them, it should be feasible to do a large scale cloning of trapped genes from the mouse ES cell lines.
Chowdhury
K
K
Department of Molecular Cell Biology, Max Planck Institute of Biophysical Chemistry, Am Fassberg, D-37077 Göttingen, Germany. kchawdh@gwdg.de
Bonaldo
P
P
Torres
M
M
Stoykova
A
A
Gruss
P
P
D000818:Animals; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D018274:Electroporation; D004798:Enzymes / Q000096:biosynthesis / Q000235:genetics; D005091:Exons; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012270:Ribosomes / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*
D004798:Enzymes; D011506:Proteins; D011994:Recombinant Proteins