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Functional gene analysis tools in are often ineffective in other nematodes due to differences in gonadal morphology and transgene silencing. Here, we established a method to generate stable transgenic lines in the nematodes and using microparticle bombardment coupled with hygromycin B selection. Despite using non-codon-optimized GFP, transgenic strains expressing fluorescent markers were obtained in both species. Additionally, an codon-optimized RFP construct showed robust expression in all tissues. This method will be valuable for future studies into the unusual sex determination, viviparity, and stress resistance in and .
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http://dx.doi.org/10.17912/micropub.biology.001585 | DOI Listing |
Curr Protoc
August 2025
Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, Iowa.
Plasmodesmata (PD) are highly specialized, nanoscopic pores that traverse the cell wall to connect the cytoplasm of adjacent plant cells, enabling direct cell-to-cell communication. PD provides the continuity of three key cellular components: the plasma membrane, the endoplasmic reticulum (ER), and the cytosol. The compressed ER within PD is known as the desmotubule.
View Article and Find Full Text PDFaBIOTECH
June 2025
Centre for Crop Science, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, 4072 Australia.
Unlabelled: The key factors for genome-editing in plants using CRISPR/Cas9, such as the Cas9 nuclease and guide RNA (gRNA) are typically expressed from a construct that is integrated into the plant genome. However, the presence of foreign DNA in the host genome causes genetic and regulatory concerns, particularly for commercialization. To address this issue, we developed an accelerated pipeline for generating transgene-free genome-edited sorghum () in the T generation.
View Article and Find Full Text PDFMicroPubl Biol
May 2025
School of Agriculture, Meiji University, Kanagawa, Japan.
Functional gene analysis tools in are often ineffective in other nematodes due to differences in gonadal morphology and transgene silencing. Here, we established a method to generate stable transgenic lines in the nematodes and using microparticle bombardment coupled with hygromycin B selection. Despite using non-codon-optimized GFP, transgenic strains expressing fluorescent markers were obtained in both species.
View Article and Find Full Text PDFFront Plant Sci
November 2023
Department of Plant Biotechnology and Bioinformatics, Ghent University, Ghent, Belgium.
The use of CRISPR/Cas9 is currently the method of choice for precise genome engineering in plants, including in the biomass crop poplar. The most commonly used method for delivering CRISPR/Cas9 and its components in poplar is via mediated transformation, that besides the desired gene-editing event also results in stable T-DNA integration. Here we explore the delivery of the gene-editing reagents via DNA-coated microparticle bombardment into the model tree to evaluate its potential for developing transgene-free, gene-edited trees, as well as its potential for integrating donor DNA at specific target sites.
View Article and Find Full Text PDFMar Biotechnol (NY)
October 2023
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, 184-8588, Koganei, Tokyo, Japan.
The oleaginous diatom Fistulifera solaris is a promising producer of biofuel owing to the high content of the lipids. A genetic transformation technique by microparticle bombardment for this diatom was already established. However, the transformation efficiency was significantly lower than those of other diatoms.
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