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Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice.
Abstract
A general strategy for selecting insertion mutations in mice has been devised. Constructs lacking a promoter and including a beta-galactosidase gene, or a reporter gene encoding a protein with both beta-galactosidase and neomycin phosphotransferase activity, were designed so that activation of the reporter gene depends on its insertion within an active transcription unit. Such insertion events create a mutation in the tagged gene and allow its expression to be followed by beta-galactosidase activity. Introduction of promoter trap constructs into embryonic stem (ES) cells by electroporation or retroviral infection has led to the derivation of transgenic lines that show a variety of beta-galactosidase expression patterns. Intercrossing of heterozygotes from 24 strains that express beta-galactosidase identified 9 strains in which homozygosity leads to an embryonic lethality. Because no overt phenotype was detected in the remaining strains, these results suggest that a substantial proportion of mammalian genes identified by this approach are not essential for development.
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The mROSA26 locus was originally identified in mouse ESCs for stable expression of promoter-less transgenes using lentivirus knockin screens.
“The mROSA26 locus was originally identified in mouse ESCs for stable expression of promoter-less transgenes using lentivirus knockin screens51”
From Exogenous expression of ATP8, a mitochondrial encoded protein, from the nucleus in vivo (2024) · cites this paper as [51]
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Metadata
- Authors
- Friedrich, G., & Soriano, P.
- Venue
- Genes & Development
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 1991-09
- Volume
- 5
- Issue
- 9
- Pages
- 1513-1523
- ISSN
- 0890-9369
- 1549-5477
- Cited by
- 1,151
- References
- 0
- Language
- en
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