Journal ArticleDOI
Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells
H te Riele,E R Maandag,E R Maandag,A Clarke,A Clarke,M Hooper,M Hooper,Anton Berns,Anton Berns +8 more
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TLDR
The feasibility of efficiently consecutive inactivation of both alleles of the pim-1 proto-oncogene in embryonic stem cells is illustrated by the efficient consecutive in activation of both allele by homologous recombination in cultured cells.Abstract:
Specific genes can be inactivated or mutated in the mouse germ line. The phenotypic consequences of the mutation can provide pivotal information on the function of the gene in development and maintenance of the mammalian organism. The procedure entails homologous recombination in embryonic stem cells, which, on fusion to recipient blastocysts, give rise to chimaeric mice that can transmit the mutant gene to their offspring. Inbreeding can then yield mice carrying the mutation in both alleles allowing the phenotypic analysis of recessive mutations. In addition to mice lacking a particular gene function, cell lines carrying null alleles of normally expressed genes can be instrumental in assessing the function of the gene. These cell lines can either be obtained from homozygous animals or, should the mutation be lethal early in embryonic development, be generated by consecutive inactivation of both alleles by homologous recombination in cultured cells. Here we illustrate the feasibility of this latter approach by the efficient consecutive inactivation of both alleles of the pim-1 proto-oncogene in embryonic stem cells.read more
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Targeted mutation of the DNA methyltransferase gene results in embryonic lethality.
TL;DR: Results indicate that while a 3-fold reduction in levels of genomic m5C has no detectable effect on the viability or proliferation of ES cells in culture, a similar reduction of DNA methylation in embryos causes abnormal development and embryonic lethality.
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Identification of DC-SIGN, a Novel Dendritic Cell–Specific ICAM-3 Receptor that Supports Primary Immune Responses
Teunis B.H. Geijtenbeek,Ruurd Torensma,S.J. van Vliet,G.C.F. van Duijnhoven,Gosse Jan Adema,Y. van Kooyk,Carl G. Figdor +6 more
TL;DR: It is demonstrated that ICAM-3 expressed by resting T cells is important in this first contact with dendritic cells, and it is predicted that DC-SIGN enables T cell receptor engagement by stabilization of the DC-T cell contact zone.
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Brca1 controls homology-directed DNA repair.
TL;DR: A caretaker role for BRCA1 is demonstrated in preserving genomic integrity by promoting homologous recombination and limiting mutagenic nonhomologous repair processes.
Journal ArticleDOI
Requirement for a functional Rb-1 gene in murine development
Alan Richard Clarke,Els C.Robanus Maandag,M van Roon,N. M. T. Van Der Lugt,M. A. Van Der Valk,Martin L. Hooper,Anton Berns,H te Riele +7 more
TL;DR: It is reported here that young heterozygous mice do not appear abnormal and do not develop retinoblastoma at a detectable frequency, however, homozygous mutant embryos fail to reach term and show a number of abnormalities in neural and haematopoietic development.
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Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding domains.
TL;DR: The construction and characterization of a series of chimeric recombinases, each consisting of Cre fused to a mutated human oestrogen receptor (ER) ligand-binding domain (LBD) are reported, which should be useful for efficient spatio-temporally controlled site-directed somatic mutagenesis.
References
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Journal ArticleDOI
Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells.
Kirk R. Thomas,Mario R. Capecchi +1 more
TL;DR: This work mutated, by gene targeting, the endogenous hypoxanthine phosphoribosyl transferase (HPRT) gene in mouse embryo-derived stem (ES) cells and compared the gene-targeting efficiencies of two classes of neor-Hprt recombinant vectors.
Journal ArticleDOI
Altering the genome by homologous recombination.
TL;DR: The current status of gene targeting with particular emphasis on germ line modification of the mouse genome is discussed, and the different methods so far employed to identify those rare embryonic stem cells in which the desired targeting event has occurred are described.
Journal ArticleDOI
At least six nucleotides preceding the AUG initiator codon enhance translation in mammalian cells
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Journal ArticleDOI
HPRT-deficient (Lesch-Nyhan) mouse embryos derived from germline colonization by cultured cells.
TL;DR: The realization of this possibility by the selection in vitro of variant ES cells deficient in hypoxanthine guanine phosphoribosyl transferase is reported, leading to their use to produce germline chimaeras resulting in female offspring heterozygous for HPRT-deficiency, and the generation of HPRt-deficient preimplantation embryos from these females.
Journal ArticleDOI
Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae
Linda Gritz,Julian Davies +1 more
TL;DR: The plasmid-borne gene hph coding for hygromycin B phosphotransferase (HPH) in Escherichia coli has been identified and its nucleotide sequence determined.
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