MASTR: A technique for mosaic mutant analysis with spatial and temporal control of recombination using conditional floxed alleles in mice Journal Article
| Authors: | Lao, Z.; Raju, G. P.; Bai, C. B.; Joyner, A. L. |
| Article Title: | MASTR: A technique for mosaic mutant analysis with spatial and temporal control of recombination using conditional floxed alleles in mice |
| Abstract: | Mosaic mutant analysis, the study of cellular defects in scattered mutant cells in a wild-type environment, is a powerful approach for identifying critical functions of genes and has been applied extensively to invertebrate model organisms. A highly versatile technique has been developed in mouse: MASTR (mosaic mutant analysis with spatial and temporal control of recombination), which utilizes the increasing number of floxed alleles and simultaneously combines conditional gene mutagenesis and cell marking for fate analysis. A targeted allele (R26 MASTR) was engineered; the allele expresses a GFPcre fusion protein following FLP-mediated recombination, which serves the dual function of deleting floxed alleles and marking mutant cells with GFP. Within 24 hr of tamoxifen administration to R26 MASTR mice carrying an inducible FlpoER transgene and a floxed allele, nearly all GFP-expressing cells have a mutant allele. The fate of single cells lacking FGF8 or SHH signaling in the developing hindbrain was analyzed using MASTR, and it was revealed that there is only a short time window when neural progenitors require FGFR1 for viability and that granule cell precursors differentiate rapidly when SMO is lost. MASTR is a powerful tool that provides cell-type-specific (spatial) and temporal marking of mosaic mutant cells and is broadly applicable to developmental, cancer, and adult stem cell studies. Mosaic mutant analysis is a powerful technique for determining cell-autonomous and nonautonomous gene functions; however, the development of approaches in mouse has lagged behind other multicellular model organisms. Joyner and colleagues describe a new technique in mouse that utilizes any floxed allele and simultaneously combines conditional gene mutagenesis and cell marking to produce mosaic mutants. MASTR is a powerful tool that provides cell-type-specific and temporal control and is broadly applicable to developmental, cancer, and adult stem cell studies. © 2012 The Authors. |
| Keywords: | signal transduction; controlled study; protein expression; unclassified drug; gene mutation; gene deletion; mutation; nonhuman; mouse; animals; mice; allele; animal tissue; cell viability; mus; gene expression; green fluorescent protein; sonic hedgehog protein; animal experiment; alleles; cell fate; neural stem cell; cell differentiation; validation study; mutational analysis; mice, transgenic; genetic recombination; hybrid protein; genetic engineering; recombination, genetic; brain development; granule cell; transgene; tamoxifen; dna mutational analysis; cre recombinase; rhombencephalon; adult stem cell; fibroblast growth factor receptor 1; cell mutant; regulator protein; invertebrata; mosaicism; chromosome disorders; fgf8 protein; green fluorescent protein cre fusion protein; floxed gene; flpoer gene; mosaic mutant analysis; mosaic mutant analysis with spatial and temporal control of recombination |
| Journal Title: | Cell Reports |
| Volume: | 2 |
| Issue: | 2 |
| ISSN: | 2211-1247 |
| Publisher: | Cell Press |
| Date Published: | 2012-08-30 |
| Start Page: | 386 |
| End Page: | 396 |
| Language: | English |
| DOI: | 10.1016/j.celrep.2012.07.004 |
| PROVIDER: | scopus |
| PMCID: | PMC3460375 |
| PUBMED: | 22884371 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 1 October 2012" - "Source: Scopus" |
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