Embryonic stem cells and mice expressing different GFP variants for multiple non-invasive reporter usage within a single animal

Background: Non-invasive autofluorescent reporters have revolutionized lineage labeling in an array of different organisms. In recent years green fluorescent protein (GFP) from the bioluminescent jellyfish Aequoria Victoria has gained popularity in mouse transgenic and gene targeting regimes 1. It offers several advantages over conventional gene-based reporters, such as lacZ and alkaline phosphatase, in that its visualization does not require a chromogenic substrate and can be realized in vivo. We have previously demonstrated the utility and developmental neutrality of enhanced green fluorescent protein (EGFP) in embryonic stem (ES) cells and mice 2. Results: In this study we have used embryonic stem (ES) cell-mediated transgenesis to test the enhanced cyan fluorescent protein (ECFP) and enhanced yellow fluorescent protein (EYFP), two mutant and spectrally distinct color variants of wild type (wt) GFP. We have also tested DsRed1, the novel red fluorescent protein reporter recently cloned from the Discostoma coral by virtue of its homology to GFP. To this end, we have established lines of ES cells together with viable and fertile mice having widespread expression of either the ECFP or EYFP GFP-variant reporters. However, we were unable to generate equivalent DsRed1 lines, suggesting that DsRed1 is not developmentally neutral or that transgene expression cannot be sustained constitutively. Balanced (diploid diploid) and polarized (tetraploid diploid) chimeras comprising combinations of the ECFP and EYFP ES cells and/or embryos, demonstrate that populations of cells expressing each individual reporter can be distinguished within a single animal. Conclusions: GFP variant reporters are unique in allowing non-invasive multi-spectral visualization in live samples. The ECFP and EYFP-expressing transgenic ES cells and mice that we have generated provide sources of cells and tissues for combinatorial, double-tagged recombination experiments, chimeras or transplantations

Assessing the Social Service Needs of an Emerging Population of Recent Mexican Immigrants Living with, or at risk for, HIV/AIDS

Latinos have recently become the largest minority group in the United States. It is expected that Latino population growth will continue due to high fertility and high immigration rates. Latinos currently experience a health disparity in HIV rates compared to Caucasians. This disparity may be associated with health care access, which is compounded by the phenomenon of an emerging population with language and culture that differ from the health care system. This article then provides an example of the methodology developed to access and assess the needs of, individuals with HIV/AIDS who traditionally have not participated in needs assessments. Implications of the current research findings and recommendations for future research are discussed.Desde hace poco tiempo la población latina se ha convertido en el grupo minoritario más grande de los Estados Unidos. Se espera que el crecimiento de este sector poblacional continúe también en el futuro debido a una elevada tasa de fertilidad y de inmigración. Comparados con la población blanca, los latinos experimentan una disparidad en cuanto a las tasas de VIH que puede estar asociada con el acceso a servicios de salud el cual se agrava ya que se trata de una población emergente con lengua y cultura diferentes a los conocidos por el sistema de salud. En este artículo se ofrecen ejemplos de una metodología que fue desarrollada para conocer y evaluar las necesidades de individuos que padecen VIH/SIDA y que tradicionalmente no han participado en este tipo de evaluaciones. Sobre esta base se discuten las implicaciones de los resultados y se elaboran una serie de recomendaciones para investigaciones futuras

Identification of genetic elements in metabolism by high-throughput mouse phenotyping

Metabolic diseases are a worldwide problem but the underlying genetic factors and their relevance to metabolic disease remain incompletely understood. Genome-wide research is needed to characterize so-far unannotated mammalian metabolic genes. Here, we generate and analyze metabolic phenotypic data of 2016 knockout mouse strains under the aegis of the International Mouse Phenotyping Consortium (IMPC) and find 974 gene knockouts with strong metabolic phenotypes. 429 of those had no previous link to metabolism and 51 genes remain functionally completely unannotated. We compared human orthologues of these uncharacterized genes in five GWAS consortia and indeed 23 candidate genes are associated with metabolic disease. We further identify common regulatory elements in promoters of candidate genes. As each regulatory element is composed of several transcription factor binding sites, our data reveal an extensive metabolic phenotype-associated network of co-regulated genes. Our systematic mouse phenotype analysis thus paves the way for full functional annotation of the genome