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Blood, 1 January 2001, Vol. 97, No. 1, pp. 324-326
BRIEF REPORT
Activation of EGFP expression by Cre-mediated excision in a new
ROSA26 reporter mouse strain
Xiaohong Mao,
Yuko Fujiwara,
Aimée Chapdelaine,
Haidi Yang, and
Stuart H. Orkin
From the 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.
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.
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