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We have developed and tested two linked but separable structured inquiry exercises using a set of Drosophila melanogaster GAL4 enhancer trap strains for an upper-level undergraduate laboratory methods course at Bucknell University. In the first, students learn to perform inverse PCR to identify the genomic location of the GAL4 insertion, using FlyBase to identify flanking sequences and the primary literature to synthesize current knowledge regarding the nearest gene. In the second, we cross each GAL4 strain to a UAS-CD8-GFP reporter strain, and students perform whole mount CNS dissection, immunohistochemistry, confocal imaging, and analysis of developmental expression patterns. We have found these exercises to be very effective in teaching the uses and limitations of PCR and antibody-based techniques as well as critical reading of the primary literature and scientific writing. Students appreciate the opportunity to apply what they learn by generating novel data of use to the wider research community.
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http://dx.doi.org/10.1371/journal.pbio.1002030 | DOI Listing |
Open Life Sci
August 2025
Biology Department, University of Pittsburg at Greensburg, Greensburg, Pennsylvania, United States of America.
Gene regulation is important during tissue formation, but redundant systems make it difficult to study The protein Jazf-1 is a member of the NuA4/TIP60 histone-modifying complex, and a transcriptional repressor has been suggested to be important for eye development. We used the GAL4-UAS system to determine the impact of altering gene expression. GAL4-UAS manipulations of Jazf-1 in the eye caused variable and not fully penetrant phenotypes.
View Article and Find Full Text PDFJ Genet Genomics
August 2025
Department of Neurology, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing 100045, China; Department of Neurology, Children's Medical Center, Peking University First Hospital, Beijing 102627, China. Electronic address:
Spinocerebellar ataxia (SCA) type 51 is a neurodegenerative disease caused by CAG repeat expansions in exon 1 of the THAP11 gene. These repeats are translated into a glutamine-rich protein, THAP11-polyQ, which forms protein aggregates and exhibits toxicity in cell models; however, the underlying mechanism remains unclear. In this study, we generate transgenic Drosophila models expressing varying lengths of THAP11-polyQ using the UAS-GAL4 system and assess neurodegeneration through pathological and behavioral analyses.
View Article and Find Full Text PDFbioRxiv
July 2025
Department of Entomology, Cornell University, Ithaca, NY, United States of America.
The ability to direct tissue-specific overexpression of transgenic proteins in genetically tractable organisms like has facilitated innumerable biological discoveries. However, transgenic proteins can themselves impact cellular and physiological processes in ways that are often ignored or poorly defined. Here we discovered that the transgene, which directs strong expression of the yeast GAL4 transcription factor in the fat body, induces significant physiological defects in adult female flies.
View Article and Find Full Text PDFPlant Cell Rep
July 2025
DOE Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
A new GAL4-based feed-forward loop circuit enhances β-glucuronidase (GUS) reporter gene expression in leaves and stems of stably transformed sugarcane plants.
View Article and Find Full Text PDFMol Ther
August 2025
Department of Life Science, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 06974, Republic of Korea. Electronic address:
Galectin-4 (Gal-4), a member of the β-galactoside-binding galectin family, plays a role in various physiological processes, including tumor progression and intestinal disorders. However, its contribution to adaptive immunity remains poorly understood. In this study, Gal-4 is identified as a critical factor for effective generation of CD8+ T cell responses against tumors and viral infections.
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