CRISPR/Cas9-based transcriptional activation (CRISPRa) has recently emerged as a powerful and scalable technique for systematic over-expression genetic analysis in Drosophila melanogaster. We present flySAM, a potent new tool for in vivo CRISPRa, which offers a major improvement over existing strategies in terms of effectiveness, scalability, and ease-of-use. flySAM outperforms existing in vivo CRISPRa strategies, and approximates phenotypes obtained using traditional Gal4-UAS over-expression. Further, because flySAM typically only requires a single sgRNA, it dramatically improves scalability. We use flySAM to demonstrate multiplexed CRISPRa, which has not been previously shown in vivo. In addition, we have simplified the experimental usage of flySAM by creating a single vector encoding both the UAS:Cas9-activator and the sgRNA, allowing for inducible CRISPRa in a single genetic cross. flySAM will thus replace previous CRISPRa strategies as the basis of our growing genome-wide transgenic over-expression resource, TRiP-OE.
High prevalence of vitamin D deficiency has been reported worldwide. Residents of Taiyuan, China, were predicted to be at high risk of vitamin D deficiency due to its high latitude, heavy air pollution, and cultural sun avoidance. This study investigated the vitamin D status of office workers, and explored the potential determinants of capillary 25-hydroxyvitamin D (25(OH)D) concentration as well as the relationship between 25(OH)D and metabolic syndrome. Two hundred participants, aged 20 to 80 years, were recruited. Capillary dried blood spot (DBS) 25(OH)D was measured; together with anthropometric (height, weight, and waist circumference), biochemical (serum lipid profile and fasting glucose) measures and a lifestyle questionnaire. Thirty-four percent of participants had 25(OH)D concentrations below 30 nmol/L, indicating deficient vitamin D status. Women’s 25(OH)D (median; 32.7 nmol/L (upper and lower quartile; 25.8, 43.8)) was significantly lower than men (44.0 nmol/L (32.3, 55.4)) (p < 0.01). Female gender, higher fasting glucose, and increased smoking (p < 0.05) were negatively associated with 25(OH)D concentration. However, there was no association found between metabolic syndrome (MetS) and 25(OH)D concentration and no significant difference in vitamin D status between men or women with MetS compared to healthy individuals. Vitamin D deficiency was common in urban residents of Taiyuan in winter and more so in women than men.
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