Activation of EGFP expression by Cre-mediated excision in a new ROSA26 reporter mouse strain.
Reporter mouse strains are important tools for monitoring Cre recombinase-mediated excision in vivo. In practice, excision may be incomplete in a given population due to threshold level or variegated expression of Cre. Hence, it is desirable in many experimental contexts to isolate cells that have undergone excision to assess the consequences of gene ablation. To generate alternative reporter mice, an enhanced green fluorescent protein (EGFP) gene was targeted to the retroviral-trapped ROSA26 locus. Upon Cre-mediated excision of "Stop" sequences, EGFP was expressed ubiquitously during embryogenesis and in adult tissues (including T cells, B cells, and myeloid cells). Using this new reporter strain, separation of excised from nonexcised cells in vitro was achieved in thymocytes in a noninvasive manner based on activated EGFP expression. This new EGFP reporter strain should facilitate a variety of conditional gene-targeting experiments, including the functional studies of hematopoietic cells in lineage-specific knockout mice.
Mao
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X
Department of Pediatrics, Division of Hematology/Oncology, Children's Hospital, the Dana Farber Cancer Institute, and the Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115, USA.
Fujiwara
Y
Y
Chapdelaine
A
A
Yang
H
H
Orkin
S H
SH
D000818:Animals; D001773:Blood Cells; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation; D018390:Gene Targeting / Q000379:methods; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D019426:Integrases / Q000378:metabolism*; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D011506:Proteins / Q000235:genetics*; D022661:RNA, Untranslated; D013950:Thymus Gland / Q000166:cytology; D014764:Viral Proteins
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011506:Proteins; D022661:RNA, Untranslated; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases