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Publication : Development of an enhanced GFP-based dual-color reporter to facilitate genetic screens for the recovery of mutations in mice.

First Author  Frank AC Year  2003
Journal  Proc Natl Acad Sci U S A Volume  100
Issue  24 Pages  14103-8
PubMed ID  14615591 Mgi Jnum  J:111135
Mgi Id  MGI:3653075 Doi  10.1073/pnas.1936166100
Citation  Frank AC, et al. (2003) Development of an enhanced GFP-based dual-color reporter to facilitate genetic screens for the recovery of mutations in mice. Proc Natl Acad Sci U S A 100(24):14103-8
abstractText  Mutagenesis screens to isolate a variety of alleles leading to null and non-null phenotypes represent an important approach for the characterization of gene function. Genetic schemes that use visible markers permit the efficient recovery of chemically induced mutations. We have developed a universal reporter system to visibly mark chromosomes for genetic screens in the mouse. The dual-color reporter is based on a single vector that drives the ubiquitous coexpression of the enhanced GFP (EGFP) spectral variants yellow and cyan. We show that widespread expression of the dual-color reporter is readily detected in embryonic stem cells, mice, and throughout developmental stages. CRE-loxP- and FLPe-FRT-mediated deletion of each color cassette demonstrates the modular design of the marker system. Random integration followed by plasmid rescue and sequence-based mapping was used to introduce the marker to a defined genomic location. Thus, single-step placement will simplify the construction of a genomewide bank of marked chromosomes. The dual-color nature of the marker permits complete identification of genetic classes of progeny as embryos or mice in classic regionally directed screens. The design also allows for more efficient and novel schemes, such as marked suppressor screens, in the mouse. The result is a versatile reporter that can be used independently or in combination with the growing sets of deletion and inversion resources to enhance the design and application of a wide variety of genetic schemes for the functional dissection of the mammalian genome.
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