Capturing genes encoding membrane and secreted proteins important for mouse development.
A strategy based on the gene trap was developed to prescreen mouse embryonic stem cells for insertional mutations in genes encoding secreted and membrane-spanning proteins. The "secretory trap" relies on capturing the N-terminal signal sequence of an endogenous gene to generate an active beta-galactosidase fusion protein. Insertions were found in a cadherin gene, an unc6-related laminin (netrin) gene, the sek receptor tyrosine kinase gene, and genes encoding two receptor-linked protein-tyrosine phosphatases, LAR and PTP kappa. Analysis of homozygous mice carrying insertions in LAR and PTP kappa showed that both genes were effectively disrupted, but neither was essential for normal embryonic development.
Skarnes
W C
WC
Biotechnology and Biological Sciences Research Council, Centre for Genome Research, University of Edinburgh, United Kingdom.
Moss
J E
JE
Hurtley
S M
SM
Beddington
R S
RS
D000818:Animals; D001483:Base Sequence; D015820:Cadherins / Q000096:biosynthesis; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D017931:DNA Primers; D004622:Embryo, Mammalian; D005314:Embryonic and Fetal Development; D005326:Fetal Proteins / Q000096:biosynthesis; D007668:Kidney; D007797:Laminin / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D017027:Protein Tyrosine Phosphatases / Q000096:biosynthesis; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis; D036122:Receptor, EphA4; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014162:Transfection; D001616:beta-Galactosidase / Q000096:biosynthesis
D015820:Cadherins; D017931:DNA Primers; D005326:Fetal Proteins; D007797:Laminin; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; D036122:Receptor, EphA4; D017027:Protein Tyrosine Phosphatases; D001616:beta-Galactosidase