Mechanisms that involve whole genome polyploidy play important roles in development and evolution; also, an abnormal generation of tetraploid cells has been associated with both the progression of cancer and the development of drug resistance. Until now, it has not been feasible to easily manipulate the ploidy of a multicellular animal without generating mostly sterile progeny. Presented here is a simple and rapid protocol for generating tetraploid Caenorhabditis elegans animals from any diploid strain. This method allows the user to create a bias in chromosome segregation during meiosis, ultimately increasing ploidy in C. elegans. This strategy relies on the transient reduction of expression of the rec-8 gene to generate diploid gametes. A rec-8 mutant produces diploid gametes that can potentially produce tetraploids upon fertilization. This tractable scheme has been used to generate tetraploid strains carrying mutations and chromosome rearrangements to gain insight into chromosomal dynamics and interactions during pairing and synapsis in meiosis. This method is efficient for generating stable tetraploid strains without genetic markers, can be applied to any diploid strain, and can be used to derive triploid C. elegans. This straightforward method is useful for investigating other fundamental biological questions relevant to genome instability, gene dosage, biological scaling, extracellular signaling, adaptation to stress, development of resistance to drugs, and mechanisms of speciation.
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http://dx.doi.org/10.3791/57296 | DOI Listing |
Adv Biotechnol (Singap)
July 2024
State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
RNA silencing (or RNA interference, RNAi) initiated by double-stranded RNAs is a conserved mechanism for regulating gene expression in eukaryotes. RNAi-based crop protection strategies, including host-induced gene silencing (HIGS), spray-induced gene silencing (SIGS) and microbe-induced gene silencing (MIGS), have been successfully used against various pests and pathogens. Here, we highlight the challenges surrounding dsRNA design, large-scale production of dsRNA and dsRNA delivery systems.
View Article and Find Full Text PDFMicroPubl Biol
January 2025
Wofford College, Spartanburg, South Carolina, United States.
Quick Oil Red O staining is a well established method to assay total lipid levels in , but the software to clean up and analyse the images is either laborious, expensive or both. We have developed a process that uses an existing protocol to stain the animals, followed by Magic Select in Paint3D to remove background and then a custom script in Biopython to quantify average pixel intensity animal. The software is free, accessible and relatively easy to use for undergraduate researchers.
View Article and Find Full Text PDFMicroPubl Biol
January 2025
Department of Neuroscience, Pomona College, Claremont, California, United States of America.
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View Article and Find Full Text PDFJ Nat Prod
January 2025
School of Pharmacy, Nantong University, 9 Seyuan Road, Nantong 226019, People's Republic of China.
Ten new resin glycosides, controlins I-X (-), were isolated from the seeds of . Their structures were established by spectroscopic analysis as well as by chemical means. Compounds were identified as glycosidic acid methyl esters, considered as artifacts generated via transesterification with MeOH from natural resin glycosides.
View Article and Find Full Text PDFPLoS Comput Biol
January 2025
European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany.
The characterization of phenotypes in cells or organisms from microscopy data largely depends on differences in the spatial distribution of image intensity. Multiple methods exist for quantifying the intensity distribution - or image texture - across objects in natural images. However, many of these texture extraction methods do not directly adapt to 3D microscopy data.
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