A novel gene trapping for identifying genes expressed under the control of specific transcription factors.
Gene trapping is a powerful method for identifying novel genes and for analyzing their functions. It is, however, difficult to select trapped genes on the basis of their function. To identify genes regulated by transcription factors that are important in the mesodermal formation, we selected trapped ES clones by infection of adenoviral vectors expressing Pax1, Brachyury, and Foxa2. Among 366 trapped genes, seven seemed to be controlled by these transcription factors in the first screening. The trapped genes were identified by 5' RACE, and a Northern blotting revealed that expressions of three trapped genes were regulated by these transcription factors. Expression patterns of Cx43 and HP1gamma implicated their functional relationships to Foxa2 in the formation of the notochord and the neural tube. Furthermore, Wtap mutant mice derived from the trapped clone showed defects in the mesendoderm formation. Our results indicate that trapped ES clones could be selected effectively using transcription factors.
Naruse
Chie
C
Division of Transgenic Animal Science, Advanced Science Research Center, Kanazawa University, Takara-machi 13-1, Kanazawa 920-8640, Japan.
Fukusumi
Yoshiyasu
Y
Kakiuchi
Dai
D
Asano
Masahide
M
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000378:metabolism; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D005822:Genetic Vectors; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008817:Mice, Mutant Strains; D016296:Mutagenesis; D009687:Nuclear Proteins / Q000235:genetics; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D000072260:RNA Splicing Factors; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D000098908:Brachyury Protein
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D005326:Fetal Proteins; D051542:Hepatocyte Nuclear Factor 3-beta; D009687:Nuclear Proteins; D051761:Paired Box Transcription Factors; D000072260:RNA Splicing Factors; D020825:T-Box Domain Proteins; D014157:Transcription Factors; D000098908:Brachyury Protein; C486008:Wtap protein, mouse; C070835:PAX1 transcription factor