Modification of Drosophila transformation vector pCaSper3 with the P element was used to construct a new vector, pFRT. The vector contains two tandem FRT sites flanked with several unique restriction sites and separated by a polylinker of five restriction sites, and allows easy cloning of DNA fragments between or close to the FRT sites. FRT-mediated excision of DNA sequences cloned between the FRT sites was demonstrated in vivo. The vector was proposed for molecular genetic studies of the position effect variegation, structural and molecular organization of Drosophila polytene chromosomes, etc.
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Proteoglycan 4 (Lubricin) and regulation of xanthine oxidase in synovial macrophage as a mechanism of controlling synovitis.
Arthritis Res Ther
December 2024
Department of Emergency Medicine, Rhode Island Hospital, Providence, RI, USA.
Background: Synovial macrophages (SMs) are important effectors of joint health and disease. A novel Cx3CR1 + TREM2 + SM population expressing the tight junction protein claudin-5, was recently discovered in synovial lining. Ablation of these SMs was associated with onset of arthritis.
View Article and Find Full Text PDFThe sparse driver system for single-cell labeling and manipulation in .
bioRxiv
December 2024
Department of Biology and Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
In this protocol, we introduce a sparse driver system for cell-type specific single-cell labeling and manipulation in , enabling complete and simultaneous expression of multiple transgenes in the same cells. The system precisely controls expression probability and sparsity via mutant sites with reduced recombination efficiency and tunable FLP levels adjusted by heat-shock durations. We demonstrate that this generalizable toolkit enables tunable sparsity, multi-color staining, single-cell trans-synaptic tracing, single-cell manipulation, and analysis of cell-autonomous gene function.
View Article and Find Full Text PDFArticle Synopsis
- Scientists found a special type of cells called synovial macrophages which help keep joints healthy and can become unhealthy in arthritis.
- They studied a protein called PRG4 which helps to keep the joints smooth and looked at how it affects these cells and inflammation using special mice that could turn off the PRG4 gene.
- Results showed that when PRG4 was turned off, it caused more inflammation and problems in the joints, but using a medicine called febuxostat helped reduce these issues and keep the healthy cells alive.
Multistep allelic conversion in mouse pre-implantation embryos by AAV vectors.
Sci Rep
August 2024
Czech Centre for Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences, Prumyslova 595, 252 50, Vestec, Czech Republic.
Site-specific recombinases (SSRs) are critical for achieving precise spatiotemporal control of engineered alleles. These enzymes play a key role in facilitating the deletion or inversion of loci flanked by recombination sites, resulting in the activation or repression of endogenous genes, selection markers or reporter elements. However, multiple recombination in complex alleles can be laborious.
View Article and Find Full Text PDFControl of ENaC ubiquitination.
Am J Physiol Renal Physiol
August 2024
Department of Physiology and Biophysics, Weill-Cornell Medicine, New York, New York, United States.
Ubiquitination influences the expression of the epithelial Na channel (ENaC). We assessed the mechanisms of selective ubiquitination of the mature, cleaved form of γENaC in both native rodent kidneys and Fisher rat thyroid (FRT) cells expressing the channel heterologously. In both models, singly cleaved and fully cleaved γENaCs were strongly ubiquitinated, implying that the second cleavage releasing an inhibitory peptide was not essential for the process.
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